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Research ArticleA Dietary Treatment Improves Cerebral Blood Flow andBrain Connectivity in Aging apoE4 Mice

Maximilian Wiesmann12 Valerio Zerbi13 Diane Jansen1 Roy Haast1 Dieter Luumltjohann4

Laus M Broersen56 Arend Heerschap3 and Amanda J Kiliaan1

1Department of Anatomy Donders Institute for Brain Cognition amp Behaviour Radboud University Medical Center6525 EZ Nijmegen Netherlands2Department of Geriatric Medicine Radboud University Medical Center 6525 EZ Nijmegen Netherlands3Department of Radiology and Nuclear Medicine Radboud University Medical Center 6525 EZ Nijmegen Netherlands4Institute for Clinical Chemistry and Clinical Pharmacology University of Bonn Bonn Germany5Nutricia Research Advanced Medical Nutrition Utrecht Netherlands6UIPS Utrecht University Utrecht Netherlands

Correspondence should be addressed to Amanda J Kiliaan amandakiliaanradboudumcnl

Received 8 January 2016 Accepted 22 February 2016

Academic Editor Tae-Jin Kim

Copyright copy 2016 Maximilian Wiesmann et al This is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in any medium provided the original work is properlycited

APOE 1205764 (apoE4) polymorphism is the main genetic determinant of sporadic Alzheimerrsquos disease (AD) A dietary approach(Fortasyn) including docosahexaenoic acid eicosapentaenoic acid uridine choline phospholipids folic acid vitamins B12 B6C and E and selenium has been proposed for dietary management of AD We hypothesize that the diet could inhibit AD-likepathologies in apoE4 mice specifically cerebrovascular and connectivity impairment Moreover we evaluated the diet effect oncerebral blood flow (CBF) functional connectivity (FC) graywhite matter integrity and postsynaptic density in aging apoE4miceAt 10ndash12 months apoE4 mice did not display prominent pathological differences compared to wild-type (WT) mice However16ndash18-month-old apoE4 mice revealed reduced CBF and accelerated synaptic loss The diet increased cortical CBF and amountof synapses and improved white matter integrity and FC in both aging apoE4 and WT mice We demonstrated that protectivemechanisms on vascular and synapse health are enhanced by Fortasyn independent of apoE genotype We further showed theefficacy of a multimodal translational approach including advanced MR neuroimaging to study dietary intervention on brainstructure and function in aging

1 Introduction

Extensive research has been pursued in search of an effectivetherapy for Alzheimerrsquos disease (AD) However no treatmentis yet available nor does it seem near Preventive approacheshave therefore consistently emerged as key policy prioritiesin recently formulated dementia strategiesThese approachesinclude modification of health-compromising behavior suchas lifestyle and dietary intake that may lead to AD Forexample the Mediterranean diet (high consumption of fruitvegetables and legumes moderate consumption of fish nutsand olive oil as the main source of fats) has been associatedwith a reduced risk of AD [1 2] and with a lower mortality

[3 4] The mechanisms via which diet influences the onsetand progression ofADpathology are still under investigationOne possible mode of action is the beneficial effect ofnutrients such as omega-3 long-chain polyunsaturated fattyacids (n3-LCPUFAs) on the vascular system [4 5]

Herewith n3-LCPUFAs will target the very early asymp-tomatic phase of the disease in which (cerebro)vascularimpairment is the strongest contributor to the onset andprogression of neurodegenerative traits of typical AD anddementia in general [6] n3-LCPUFAs may diminish severityof vascular risk factors like atherosclerosis [7] high bloodpressure [8] and other cardiovascular diseases [7 9ndash11]which are risk factors for AD as well Other nutrients may

Hindawi Publishing CorporationNeural PlasticityVolume 2016 Article ID 6846721 15 pageshttpdxdoiorg10115520166846721

2 Neural Plasticity

instead directly protect synaptic integrity For instance theformation of phosphatidylcholine the most common phos-phatide in the brain and a major component of the synapticmembrane is enhanced due to presence of its precursors inthe diet [12 13] Several preclinical studies confirmed thesefindings showing that animals supplemented with the com-bination of these membrane precursors showed increasedlevels of brain phospholipids dendritic spines and neuriteoutgrowth with beneficial effects on cognition [14ndash18] Basedon these findings a novel multinutrient supplementation dietcalled Fortasyn comprising n3-LCPUFAs docosahexaenoicacid (DHA) and eicosapentaenoic acid (EPA) besides otherprecursors and cofactors for membrane synthesis such asuridine choline phospholipids folic acid vitamins B12 B6C and E and selenium has been proposed for the dietarymanagement of AD [19] To date two randomized controlledclinical trials have shown improvements in the delayed verbalrecall task and better cognitive performance in mild ADpatients supplemented with this nutrient combination [20ndash22] Although it has been recognized that Fortasyn addressesspecific nutritional needs in early AD and that it improvesfunctional connectivity as assessed by EEG [23] other pro-cesses by which Fortasyn may influence the pathophysiologyof AD need to be further elucidated and more studies arerequired to confirm its efficacy

Some studies suggested an interaction of the cholesteroltransporter apolipoprotein (apoE) with lipid-based dietaryintervention [24ndash26] ApoE is a 34 kDa glycoprotein thatexists in three isoforms apoE-1205762 apoE-1205763 and apoE-1205764which differ by one or two amino acid residues 112 and 158[27] This small difference strongly affects the conformationand the structure of apoE and influences its ability to bindlipids receptors and amyloid-120573 (A120573) [28] ApoE polymor-phic alleles have been identified as the main genetic deter-minants of AD risk specifically apoE-1205764 has been associatedwith increased toxicity and loss of neuroprotective functionin the pathogenesis of Alzheimerrsquos disease dependent on orindependent of A120573 accumulation [29] Importantly someof these processes are directly or indirectly linked to animpaired vascular system [30ndash32] The development of anappropriate animal model targeting the murine APOE genefor replacement with the human APOE-1205764 (apoE-1205764apoE4mouse) [33] opened a window for new possibilities of char-acterizing the apoE-1205764 phenotype and of studying the effectsof very early AD-like pathology development in relation withlipid-based treatment ApoE4 carrier mice exhibit an alteredlipid profile with increased risk of atherosclerosis plaquesformation [33] Altered behavior and cognitive deficits havealso been reported [34] A previous study from our groupshowed that a long-term dietary intervention with the For-tasyn diet was able to reduce anxiety behavior in 10-month-old apoE4 mice [35] Transgenic AD mice on the same dietshowed restored cortical cerebral blood flow (CBF) and brainstructural integrity compared to wild-type mice [17]

Following these promising findings we hypothesize thata nutritional intervention with Fortasyn may be able torescue or prevent the occurrence of early AD-like pathologiesin apoE4 mice such as cerebrovascular impairment andconcomitant brain connectivity loss To test this hypothesis

we evaluated the effect of the Fortasyn diet on cerebraland plasma levels of fatty acids and sterols cerebral bloodflow (CBF) gray and white matter integrity functionalconnectivity (FC) and postsynaptic density during aging in12- and 18-month-old apoE4 mice

2 Materials and Methods

21 Animals The apoE-1205764 founder mice were originallyobtained from Taconic Transgenic Models (Hudson NYUSA) and a colonywas established at the RadboudUniversityMedical Center (Radboudumc) ApoE4 mice were createdby targeting the murine apoE gene for replacement with thehuman apoE-1205764 alleles cultured in E14TG2a embryonic stem(ES) cells as described previously [36] Resulting chimeraswere backcrossed to C57BL6J mice for 8 generations Theline was derived by embryo transfer and is maintained byinbreeding homozygous mice For the present study maleand female apoE4 breeder mice were used to generatehomozygous apoE4 offspring (3rd generation)

C57BL6J wild-type (WT) mice obtained from ourcolony at the Radboudumc were used as controls Through-out the experiment animals were housed in groups of 2ndash7mice per cage in a controlled environment homogenouslyilluminated by normal fluorescent room light at 60 lux withroom temperature at 21∘C and an artificial 12 12 h light darkcycle (lights on at 7 am) Food and water were available adlibitum

The experiments were performed according to Dutchfederal regulations for animal protection The VeterinaryAuthority of the Radboudumc Netherlands approved all theprotocols within this study (RU-DEC 2011-058)

22 Diets and Timeline of Experimental Design Starting at2 months of age mice were randomly divided into twogroups animals were fed either with control diet or withFortasyn diet that differed with respect to the presence ofa specific combination of dietary precursors and cofactorsthat is uridine docosahexaenoic acid eicosapentaenoic acidcholine phospholipids folic acid vitamins B12 B6 C andE and selenium (Table 1) Both diets were isocaloric andbased on AIN-93M [37] with 5 fat The control dietcontained 19 soy oil 09 coconut oil and 22 corn oilthe Fortasyn diet contained 01 coconut oil 19 corn oiland 30 fish oil The Fortasyn-based diet contains a specificmultinutrient composition comprising nucleotides omega-3PUFAs choline B vitamins phospholipids and antioxidants(Table 1) Diets were manufactured and pelleted by Ssniff(Soest Germany) and stored at minus20∘C until use The firstgroup of mice underwent MR imaging at 11 (average age106 plusmn 01 months) to 12 (average age 123 plusmn 01 months)months of age and was sacrificed immediately thereafterThesecond group was scanned at 16 months of age (average age162plusmn01months) and sacrificed at 18 months of age (averageage 179 plusmn 01 months) The timeline of the experimentaldesign is illustrated in Figure 1 The sample size of minimal6 mice (12-month WT-control 119899 = 9 WT-Fortasyn 119899 = 9apoE4-control 119899 = 8 and apoE4-Fortasyn 119899 = 10 18-monthWT-control 119899 = 10 WT-Fortasyn 119899 = 10 apoE4-control

Neural Plasticity 3

Table 1 Compositions of the experimental diets used based onAIN-93M [37] with minor revisions

SourceDietary groups

Control[g100 g]

Fortasyn[g100 g]

Corn starch 3557 3312Casein 1400 1400Corn dextrin 155 1550Sucrose 1000 1000Dextrose 1000 1000Fibers 500 500Mineral mix (AIN-93 M-MX) 350 350Vitamin mix (AIN-93-VX) 100 100FatsSoy oil 1900 mdashCoconut oil 0900 0100Corn oil 2200 1870Fish oil mdash 3030Additionsl-Cysteine 0180 0180Choline bitartrate (411 choline) 0250 0250tert-Butylhydroquinone 00008 00008PyridoxinendashHCL mdash 000328Folic acid (90) mdash 000067Cyanocobalamin (01 in mannitol) mdash 000350Ascorbic acid (100 pure) mdash 0160dl-120572-Tocopheryl acetate (500 IUg) mdash 04650UMP disodium (24 H

2

O) mdash 10Choline chloride (74576) mdash 0402Soy lecithin mdash 0402Sodium selenite (46min) mdash 000023Energy (kcal100 g chow) 3769 3671

BirthStart diets

0 2 4 10 12 14 16 18

MRI amp dagger MRI amp dagger

Months

Figure 1 Timeline of experimental design Starting from 2 monthsof age mice were randomly divided into two groups animals werefed either with control diet or with Fortasyn diet The first group ofmice underwent MR imaging (MRI) at 11 to 12 months of age andwere sacrificed (dagger) immediately thereafter The second group wassacrificed after MRI at 18 months of age

119899 = 10 and apoE4-Fortasyn 119899 = 6) per group was chosenbased on formal calculation of power as described in theapproved protocols (RU-DEC 2011-058)

23 MR Imaging MRI measurements were performed onan 117 T BioSpec Avance III small animal MR system

(Bruker BioSpin Ettlingen Germany) equipped with anactively shielded gradient set of 600mTm and operated byParaVision 51 software between 8 am and 8 pm We useda circular polarized volume resonator for signal transmis-sion and an actively decoupled mouse brain quadraturesurface coil for signal reception (Bruker BioSpin) Duringthe MR experiments low-dose isoflurane was used (35for induction and sim15 for maintenance) slightly adjustedthroughout the experiment to maintain a fast and stablebreathing frequency (gt130 bpm) The mice were placed in astereotactic device in order to immobilize the head Bodytemperature was measured with a rectal thermometer andmaintained at 37∘C by a heated air flow device

After standard adjustments and shimming gradient echo(GE) T

2

lowast-weighted images covering the entire mouse brainwere acquired for anatomical reference

To study brain perfusion under resting conditions weused a flow-sensitive alternating inversion recovery arterialspin labelling (FAIR ASL) technique [17 38] Briefly fif-teen images with increasing inversion times (TIs) (40msndash3000ms) were obtained for the T

1calculations amounting to

a total scan time of 12 minutes Inversion recovery data fromthe imaging slice were acquired after selective inversion inter-leaved with nonselective inversion Relative cerebral bloodflow (CBF) was measured in the cortex in the hippocampusand in the thalamus based on [17]

Diffusion of water was imaged as described previously[35 39] In short 22 axial slices covering the whole brainwere acquired with a four-shot SE-EPI protocol B0 shiftcompensation navigator echoes and an automatic correctionalgorithm to limit the occurrence of ghosts and artefacts wereimplemented Encoding b-factors of 0 smm2 (b0 images5x) and 1000 smm2 were used and diffusion-sensitizinggradients were applied along 30 noncollinear directions inthree-dimensional space

The diffusion tensor was estimated for every voxel usingthe PATCH algorithm [40] mean water diffusivity (MD)and fractional anisotropy (FA) were derived from the tensorestimation following a protocol as described elsewhere [35]MD and FA values were measured in several white matter(WM) and gray matter (GM) areas which were manuallyselected based on an anatomical atlas [41]

The resting-state fMRI (rsfMRI) datasets were firstrealigned using a least-squares method and rigid-body trans-formation with Statistical Parametric Mapping (SPM) mousetoolbox (SPM5 University College London httpwwwfilionuclacukspm [42])Mean andmaximumdisplacementacross the six degrees of freedom (along the 119909- 119910- and 119911-axes and on three rotation parameters pitch roll and yaw)were measured in each mouse The mean SE-EPI imagesof each mouse were then used to generate a study-specifictemplate through linear affine and nonlinear diffeomorphictransformation (ANTs v19 httppicslupenneduANTS)Visual inspection of the normalized dataset was performedto check for possible normalization biases On the template15 areas were selected in left and right hemisphere Theselected regions were based on previous work in functionalconnectivity in mice [43] and included left and right dorsal

4 Neural Plasticity

hippocampus left and right ventral hippocampus left andright auditory cortex left and right motor cortex left andright somatosensory cortex and left and right visual cortexAll cortical ROIs were selected 1-2 voxels away from theedge of the cortex to minimize the impact of susceptibility-weighted artefacts which are more prominent in areas ofdifferent tissues interface (eg near the skull or near the earcanals) After motion regression in-plane spatial smoothing(04 times 04mm) linear detrending and temporal high-passfiltering (cutoff at 001Hz) were applied to compensatefor small across-mouse misregistration and temporal low-frequency noise FC group comparison between ROIs wascalculated from the BOLD time series using total cor-relation and partial correlation analyses implemented inFSLNets (FSLNets v03 httpfslfmriboxacukfslfslwiki)Pearsonrsquos correlation values were Fisher transformed to 119885-scores for group comparisons and statistical analysis

24 Brain Tissue Preparation Directly following the MRmeasurements at 12 and 18 months of age anaesthetisedmice were sacrificed by transcardial perfusion with 01Mphosphate buffered saline (PBS) The perfused brains werecut sagittally and the right hemispheres were snap frozenin liquid nitrogen and stored at minus80∘C before further bio-chemical processing The left hemispheres were immersionfixated for 15 h at 4∘C in 4 paraformaldehyde fixative andthereafter stored in 01M PBSwith 001 sodium azide at 4∘Cfor immunohistochemical staining

25 Immunohistochemistry PSD95 Eight series of 30 120583mcoronal sections were cut through the brain using a slid-ing microtome (Microm HM 440 E Walldorf Germany)equipped with an object table for freeze sectioning at minus60∘CThe tissue was stained for postsynaptic density with poly-clonal rabbit anti-PSD95 antibody (1 2000 Abcam Cat ab18258 RRIDAB 444362) using one complete series ofbrain sections Immunohistochemistry was performed usingstandard free-floating labelling procedures as described pre-viously

26 Quantification The stained sections were analysed usinga Zeiss Axioskop microscope equipped with hardware andsoftware of Microbrightfield (Williston VT USA) Brainregions were based on the mouse brain atlas of Franklin andPaxinos (third edition 2008) and quantified in five regionsof the hippocampus the inner molecular layer (IML) outermolecular layer (OML) cornus ammonis 1 (CA1) CA2 andCA3 Additionally two regions in the cortex correspondingto the visual and somatosensory cortex were analysed Therelevant regions were digitized at 100x magnification withimmersion oil using Stereo Investigator The quantificationof the photographs was performed using Image J (Image JUS National Institutes ofHealth BethesdaMarylandUSA)The contrast was manually enhanced following the sameprocedure for all digitized images and the amount of tissuestained was measured with a threshold-based approach

27 Biochemical Analyses Serum and brain sterol levelswere measured by gas-chromatography-mass-spectrometry-selected-ion-monitoring (GC-MS-SIM) as described in detailpreviously [44] The cerebellum of the right hemispherewas homogenized and sterols were extracted overnight bychloroformmethanol trimethylsilylation prior to GC-MS-SIM analysis [44] Brain fatty acid analyses were performedwith a part of the brain homogenate (olfactory bulb and partof frontal cortex) as described previously [35]

28 Statistics For the statistical analysis IBM SPSS 20 soft-ware (IBMCorporationNewYorkNYUSA)was used Sincethe setup of the current study was designed to determine theeffects of diet supply at two stages in which apoE4 mice maydevelop different neuropathological traits of AD statisticalanalyses were performed separately for the two age-points

Multivariate ANOVA (MANOVA) with Bonferroni cor-rections using body weight as covariate when necessary wasconducted with between-group-factors genotype and diet toanalyse possible differences in all the other parameters If theBonferroni post hoc test indicated a significant interactionbetween genotype and diet the data were split for theconcerning factor and thereafter analysed again with theMANOVA Statistical significance was set at 119901 le 005 Alldata are expressed as mean plusmn SEM

3 Results

Mortality rate in the apoE4 mice was normal until age of 18months (lt8 total mortality rate) in both diet groups Noneof the WT animals died during the experiment

31 Body Weight

10ndash12-Month-Old Mice Body weight was measured at 10 and12 months of age (Figure 2(a)) Statistical analysis revealed asignificant genotype times diet interaction (119901 = 0050) ApoE4andWTmice fed the Fortasyn diet were significantly heavierthan animals on control diet (ApoE4 119865(1 18) = 466 119901 lt0001 WT 119865(1 17) = 312 119901 lt 0001) ApoE4 mice onFortasyn were significantly heavier compared toWTmice onFortasyn (119865(1 17) = 73 119901 = 0015)

At the start of the MRI measurements a significantgenotype times diet interaction on body weight was found(119901 = 0018) Again all animals on Fortasyn diet showed anincreased body weight compared to animals on control diet(ApoE4 119865(1 18) = 501 119901 lt 0001 wild-type 119865(1 17) = 67119901 = 0019) ApoE4 mice on Fortasyn were heavier than wild-type mice on the same diet (119865(1 17) = 76 119901 = 0013)

16ndash18-Month-Old Mice Body weight was measured at 16 and18months of age (Figure 2(b)) In bothmeasurements ApoE4mice weighed significantly less than WT mice (16 months119865(1 33) = 103 119901 = 0003 18 months 119865(1 33) = 90119901 = 0005) Both at 16 and at 18 months of age all animalson Fortasyn were heavier than animals on control diet (16months 119865(1 33) = 56 119901 = 0024 18 months 119865(1 33) = 45119901 = 0042)

Neural Plasticity 5

ControlFortasyn

lowast

lowastlowast

lowastlowastlowastlowastlowastlowast

lowastlowastlowast

ApoE4Wild-type10-month


0

10

20

30

40

50

60

70Bo

dy w

eigh

t (g)

(a)

ControlFortasyn

lowast

lowastlowast

lowast

lowastlowast

Wild-type ApoE416-month


0

10

20

30

40

50

60

70

Body

wei

ght (

g)

(b)

Figure 2 Body weight was measured in 10ndash12-month-old (a) and 16ndash18-month-old ApoE4 and wild-type (WT) mice (a) At 10 months ofage ApoE4 and also wild-type mice on Fortasyn were significantly heavier than on control diet (ApoE4 119901 lt 0001 wild-type 119901 lt 0001)Only on Fortasyn diet was a genotype effect found showing a higher weight of ApoE4mice compared with wild-type mice (119901 = 0015) Againat 12 months of age all animals on Fortasyn diet showed an increased body weight compared to animals on control diet (ApoE4 119901 lt 0001wild-type 119901 = 0019) ApoE4 mice on Fortasyn were heavier than wild-type mice on the same diet (119901 = 0013) (b) At 16 and also 18 monthsof age ApoE4 mice were significantly lighter than wild-type mice (16-month 119901 = 0003 18-month 119901 = 0005) Furthermore all animals onFortasyn were heavier than animals on control diet (16-month 119901 = 0024 18-month 119901 = 0042) lowast119901 le 005 lowastlowast119901 le 001 lowastlowastlowast119901 le 0001

32 Magnetic Resonance Imaging

321 Cerebral Blood Flow To study group-related differenceson cerebrovascular health we measured cerebral blood flow(CBF) with a flow-sensitive MRI technique (FAIR ASL)Three regions of interest (ROIs) on the left and right brainhemispheres were analysed cortex hippocampus and tha-lamus Since no intraindividual differences in CBF betweenright and left hemispheres were detected between micegroups (data not shown) values from both sides were aver-aged In all measures CBF was not significantly influencedby body weight

In the 12-month-old mice we detected a genotype times dietinteraction in the thalamus (119901 = 0045) In detail Fortasyndiet increased thalamic CBF (119865(1 16) = 50 119901 = 0040)more strongly in WT mice than in the apoE4 littermates(Figure 3(a))

In the 18-month-old mice (Figure 3(b)) CBF wasdecreased in the cortex (119865(1 31) = 44 119901 = 0044) and inthe thalamus (119865(1 31) = 57 119901 = 0023) of apoE4 mice ascompared to WT mice Cortical CBF was increased by theFortasyn diet in both WT and apoE4 mice (119865(1 31) = 47119901 = 0038)

322 Diffusion Tensor Imaging Quantitative assessment ofdiffusion tensor derived indices was performed for ROIsdrawn in white and gray matter regions to assess genotypeand diet effects in apoE4 and nontransgenic WT mice(Figure 4(a))

Fractional Anisotropy In the 12-month-old mice we did notdetect any significant differences in white matter FA betweenthe groups of mice (Figure 4(b))

At 18 months of age (Figure 4(c)) Fortasyn fed miceshowed a lower FA at minus07mm in the corpus callosumcompared to control fed mice (119865(1 31) = 47 119901 = 0008)

Mean Diffusivity In the 12-month-old mice the MANOVArevealed a genotype times diet interaction for MD in the motorcortex (119901 = 0024 Figure 4(b))

In detailWTmice on Fortasyn diet had higherMD in themotor cortex than wild-type mice on control diet (119865(1 14) =92 119901 = 0009) Furthermore apoE4 mice on Fortasyn diethad a lower MD in the motor cortex than WT mice onFortasyn (119865(1 17) = 52 119901 = 0036) Fortasyn diet decreasedMD in the auditory cortex (119865(1 29) = 49 119901 = 0034) andin the somatosensory cortex (119865(1 29) = 79 119901 = 0009)compared to control diet irrespective of genotype

At 18 months of age (Figure 4(c)) apoE4 mice displayedan increased MD in the auditory cortex (119865(1 29) = 56 119901 =0025) compared to WT mice

323 rsfMRI

Total Correlation Analyses To compare the FC patternsbetween different genotypes and diets rsfMRI data werestatistically analysed based on total correlation (Figure 5) andpartial correlation (Figure 6)

At 12 months of age multivariate ANOVA (MANOVA)revealed some significant genotype but no diet effects inapoE4 compared to WT mice More specifically apoE4 miceshowed reduced FC between the right auditory cortex and theleft dorsal hippocampus (119865(1 24) = 51 119901 = 0033) and alsobetween the left visual cortex and the right auditory cortex(119865(1 24) = 55 119901 = 0028)

At 18 months of age theMANOVA demonstrated severalsignificant genotype and diet effects In detail apoE4 mice

6 Neural Plasticity

12-month-oldlowast

0

100

200

300

Cortex Hippocampus Thalamus

ApoE4 ApoE4 ApoE4Wild-type

Wild-type

Wild-type

CBF

(mL100

gm

in)

ControlFortasyn

(a)

0

100

200

300


18-month-old

lowast lowast

lowast


Wild-type

Wild-type

CBF

(mL100

gm

in)

ControlFortasyn

lowast

(b)

Figure 3 Cerebral blood flow (CBF) as measured with a flow-sensitive MRI technique (FAIR ASL) at 12 months and 18 months of age inC57BL6J wild-type control mice and ApoE4 transgenic mice on control diet or Fortasyn diet Three regions of interest (ROIs) on the leftand right brain hemispheres were analysed cortex hippocampus and thalamus (a) In the 12-month-old mice we detected a genotype times dietinteraction in the thalamus (119901 = 0045) Fortasyn diet increased thalamic CBF (119901 = 0040) more strongly in wild-type mice than in the apoE4littermates (b) In the 18-month-old mice CBF was decreased in the cortex (119901 = 0044) and in the thalamus (119901 = 0023) of apoE4 mice ascompared to wild-type mice Cortical CBF was increased by the Fortasyn diet in both wild-type and apoE4 mice (119901 = 0038) lowast119901 le 005lowastlowast

119901 le 001 lowastlowastlowast119901 le 0001

had significant lower FC between cortical and hippocampalregions but also between cortical regions themselvesNotablyFortasyn was able to increase the hippocampal FC andalso FC between the visual and retrosplenial cortex to thehippocampus

The MANOVA revealed genotype times diet interactions inthe left auditory cortex to retrosplenial cortex (119901 = 0011) inthe right auditory cortex to right somatosensory cortex (119901 =005) and in the left somatosensory cortex to retrosplenialcortex (119901 = 0025) ApoE4 mice on control diet showed areduced FC between left auditory cortex and retrosplenialcortex (119865(1 15) = 179 119901 = 0001) right auditory cortex andright somatosensory cortex (119865(1 15) = 49 119901 = 0042) andleft somatosensory cortex and retrosplenial cortex (119865(1 15) =182 119901 = 0001) which were not observed in ApoE4 mice onFortasyn diet

Moreover compared to their transgenic littermates oncontrol diet only apoE4 mice on Fortasyn displayed anincreased FC between left auditory cortex and retrosplenialcortex (119865(1 11) = 130 119901 = 0004) right auditory cortex andright somatosensory cortex (119865(1 11) = 99 119901 = 0009) andleft somatosensory cortex and retrosplenial cortex (119865(1 11) =102 119901 = 0009)

Partial Correlation Analyses At 12 months of age significantgenotype and diet effects were shown using MANOVA(Figure 6) In detail apoE4 mice showed a reduced FCbetween left dorsal and ventral hippocampus (119865(1 24) = 47119901 = 0040) Additionally animals on Fortasyn diet showed anincreased partial correlation in the interhemispheric connec-tion between left and right ventral hippocampus (119865(1 24) =73 119901 = 0012)

At 18 months of age the MANOVA showed severalsignificant genotype and diet effects (Figure 6) ApoE4 micehad a reduced FC between the right auditory cortex andthe right motor cortex (119865(1 26) = 164 119901 lt 0001) TheFortasyn diet increased FC between left and right motorcortices (119865(1 26) = 71 119901 = 0013) but slightly reduced FCbetween left and right dorsal hippocampus (119865(1 26) = 44119901 = 0045)

MANOVA also revealed a genotype times diet interactionbetween right auditory cortex and right somatosensory cor-tex 119901 = 0010 Fortasyn diet increased partial correlation inapoE4 mice compared to WT between right auditory cortexand right somatosensory cortex (119865(1 12) = 81 119901 = 0015)

33 Postsynaptic Density-95 (PSD-95) Protein Density ofpostsynaptic density proteins was visualised and quantifiedimmunohistochemically in various cortical and hippocampalareas with polyclonal rabbit anti-PSD-95 At 12 monthswe did not observe significant differences between groups(Figure 7) At 18 months reduced PSD-95 levels were seenin the sensory cortex of apoE4 mice compared to WT mice(119865(1 33) = 58 119901 lt 0021) Fortasyn diet increased levels ofPSD-95 in the sensory cortex (119865(1 33) = 67119901 lt 0014) CA3area (119865(1 31) = 99 119901 lt 0004) and IML (119865(1 31) = 99119901 lt 0004) irrespective of genotype No genotype times dietinteractions were observed

34 Fatty Acids Fatty acid content was analysed in the brainsof apoE4 and WT mice (Supplementary Material + Supple-mentary Table 1 available online at httpdxdoiorg10115520166846721) At both 12 and 18 months of age increased

Neural Plasticity 7FA

MD

+03mm minus02mm minus07mm minus12mm minus27mmminus22mmminus17mm(a)

12-month-old

ApoE

4W

ild-ty

pe

tractOptic

Wild

-type

ApoE

4

CCminus07mm

Wild

-type

ApoE

4

CCminus17mm

ApoE

4W

ild-ty

pe

CCminus22mm

ApoE

4W

ild-ty

pe

Fornix

02

04

06

08

Frac

tiona

l ani

sotro

py (F

A)

lowastlowast

ControlFortasyn

lowast

lowastlowast

00006

00007

00008

00009

00010

Mea

n di

ffusiv

ity (M

D) (

mm

2s

)

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

Motorcortex

Auditorycortex

Sensorycortex

Visualcortex

ApoE

4W

ild-ty

pe

Hippocampus

(b)

ControlFortasyn

lowastlowast

00006

00007

00008

00009

00010

Mea

n di

ffusiv

ity (M

D) (

mm

2s

)

Wild

-type

ApoE

4

Motorcortex

Wild

-type

ApoE

4

Auditorycortex

Wild

-type

ApoE

4

Sensorycortex

Wild

-type

ApoE

4Visualcortex

Wild

-type

ApoE

4

Hippocampus

18-month-old

lowastlowast

ApoE

4W

ild-ty

pe

Fornix

Wild

-type

ApoE

4

CCminus22mm

Wild

-type

ApoE

4

CCminus17mm

Wild

-type

ApoE

4

CCminus07mm

Wild

-type

ApoE

4

tractOptic

02

04

06

08

Frac

tiona

l ani

sotro

py (F

A)

(c)

Figure 4 Quantitatively assessed diffusion tensor derived indices at 12 months and 18 months of age in C57BL6J wild-type control mice andApoE4 transgenic mice on control diet or Fortasyn diet (a) Fractional anisotropy (FA) and mean diffusivity (MD) were measured for ROIsdrawn in white and gray matter regions respectively (b) In 12-month-old mice no differences in FA were observed In the 12-month-oldmice the wild-type mice on Fortasyn diet had higher MD in the motor cortex than wild-type mice on control diet (119901 = 0009) FurthermoreapoE4 mice on Fortasyn diet had a lower MD in the motor cortex than wild-type mice on Fortasyn (119901 = 0036) Fortasyn diet decreased MDin the auditory cortex (119901 = 0034) and in the somatosensory cortex (119901 = 0009) compared to control diet irrespective of genotype (c) At 18months of age Fortasyn fed mice had a lower FA at minus07mm in the corpus callosum than control fed mice (119901 = 0008) At 18 months of ageapoE4 mice displayed an increased MD in the auditory cortex (119901 = 0025) lowast119901 le 005 lowastlowast119901 le 001 lowastlowastlowast119901 le 0001

relative levels of omega-3 fatty acids (119901 = 0000) and anincreased ratio of omega 36 were found in Fortasyn fedmicecompared to their littermates on control diet (119901 = 0000)At 18 months of age ApoE4 mice displayed significantlyincreased relative arachidonic acid (119901 = 0022) and relativeomega-6 content (119901 = 0038) compared to wild-type mice

For a detailed overview see the fatty acid section in theSupplementary Material and Supplementary Table 1

35 Sterol Levels Sterol levels were determined in the bloodplasma (serum) and in the cerebellum of the brain Themain findings are described below for all detailed results see

8 Neural Plasticity

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

Fortasyn

Control

Fortasyn

ApoE

4

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex

Retrosplenial cortex

Dorsal Hipp


Motor cortex

SS cortex

Visual cortex

Dorsal Hipp

Ventral Hipp

Auditory cortex

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp

12-month-oldControl

Wild

-type

Retrosplenial cortexVisual cortex

Ventral Hipp

Auditory cortex

Motor cortex

Dorsal Hipp

SS cortex

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

Wild

-type

ApoE

4

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

Fortasyn

Control

Fortasyn

18-month-oldControl

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Motor cortex

SS cortex

Visual cortex

Dorsal Hipp

Ventral Hipp

Auditory cortex

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


SS cortex

Visual cortex

Ventral Hipp

Auditory cortex

Motor cortex

Dorsal Hipp

ApoE

4W

ild-ty

peW

ild-ty

peAp

oE4

Figure 5 Resting-state functional connectivity (FC) based on total correlation analyses of 15 regions of interest (ROIs) in the mouse brainTotal correlation matrices of wild-type and apoE4 at 12 and 18 months of age both on Fortasyn and control diets At 12 months of agemultivariate ANOVA (MANOVA) revealed some significant genotype but no diet effects in apoE4 compared to wild-type mice Morespecifically apoE4 mice showed reduced FC between the right auditory cortex and the left dorsal hippocampus (119901 = 0033) and also betweenthe left visual cortex and the right auditory cortex (119901 = 0028) At 18 months of age apoE4 mice had significant lower FC between corticaland hippocampal regions but also between cortical regions themselves Notably Fortasyn was able to increase the hippocampal FC and alsoFC between the visual and retrosplenial cortices to the hippocampus

Supplementary Material and Supplementary Tables 2 and 3respectively

351 Blood Serum At 12 months of age apoE4 mice dis-played increased level of the cholesterol precursor dihy-drolanosterol (119901 = 0000) compared to WT mice At 12

months of age apoE4mice on control diet had higher levels oflathosterol (119901 = 0002) and lanosterol (119901 = 0003) comparedto WT mice on control diet

At 18 months of age apoE4 and WT mice on controldiet displayed increased levels of campesterol (119901 = 0000)sitosterol (119901 = 0016) lanosterol (119901 = 0010) desmosterol

Neural Plasticity 9

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

12-month-old

Control

Fortasyn

Control

Fortasyn

ApoE

4W

ild-ty

pe

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Dorsal Hipp

Motor cortex

SS cortex

Visual cortex

Ventral Hipp

Auditory cortex


Ventral Hipp

Auditory cortex

Motor cortex

Visual cortex

Dorsal Hipp

SS cortex

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

Wild

-type

ApoE

4

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

18-month-old

Control

Fortasyn

Control

Fortasyn

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Dorsal Hipp

Motor cortex

SS cortex

Visual cortex

Ventral Hipp

Auditory cortex


Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex

Dorsal Hipp

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp

ApoE

4W

ild-ty

peW

ild-ty

peAp

oE4

Figure 6 Resting-state functional connectivity (FC) based on partial correlation analyses of 15 regions of interest (ROIs) in the mouse brainTotal correlation matrices of wild-type and apoE4 at 12 and 18 months of age both on Fortasyn and control diets At 12 months of age apoE4mice showed a reduced FC pattern between left dorsal and ventral hippocampus (119901 = 0040) Additionally animals on Fortasyn comparedto control diet showed an increased partial correlation in the interhemispheric connection between left and right ventral hippocampus (119901 =0012) At 18 months of age ApoE4 mice on both diets had a reduced FC between the right auditory cortex and the right motor cortex(119901 lt 0001) The Fortasyn diet caused contrasting effects a higher FC was found between left-to-right motor cortices (119901 = 0013) but aslight FC reduction was revealed between left and right dorsal hippocampus (119901 = 0045) Fortasyn diet increased partial correlation in apoE4mice compared to their wild-type littermates between left and right auditory cortex (119901 = 0015) and between right auditory cortex and rightsomatosensory cortex (119901 = 0015) In wild-type mice FC between left somatosensory cortex and left auditory cortex (119901 = 0041) was higherin Fortasyn fed animals compared to control fed animals

10 Neural Plasticity

12-month-old

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

Visualcortex

Sensorycortex

IML(DG)(DG)

OMLCA1area (SR)area (SL)

CA3

0

50000

100000

150000

PSD

-95

stai

ned

area

ControlFortasyn

(a)

lowast lowastlowast

lowastlowast

lowast

lowast

18-month-old

Visualcortex

Sensorycortex

IML(DG)(DG)


CA3

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

0

50000

100000

150000

PSD

-95

stain

ed ar

ea

ControlFortasyn

(b)

Figure 7 Levels of postsynaptic density-95 (PSD-95) in various brain areas in wild-type mice and apoE4 mice on either control diet orFortasyn diet (a) At 12 months we did not find significant differences between groups (b) At 18 months reduced PSD-95 levels were seenin the sensory cortex of apoE4 mice compared to wild-type mice (119901 = 0021) Fortasyn diet increased levels of PSD-95 in the sensory cortex(119901 = 0014) CA3 area (119901 = 0004) and IML (119901 = 0004) irrespective of genotype No genotype times diet interactions were observed lowast119901 le 005lowastlowast


(119901 = 0000) 24OH-cholesterol (119901 = 0002) and cholesterol(119901 = 0033) compared to apoE4 and WT mice on Fortasyndiet

352 Cerebellum At 12 months of age apoE4 and WT miceon control diet demonstrated increased levels of lathosterol(119901 = 0003) campesterol (119901 lt 0000) and lanosterol (119901 =0001) in the cerebellum

At 18 months of age cerebellar cholesterol levels wereunchanged in ApoE4 mice compared to WT mice (119901 =0614) In control fed mice levels of campesterol and pre-cursors of cholesterol lathosterol and lanosterol were higherthan in Fortasyn fed mice (campesterol 119901 lt 0000 lathos-terol 119901 lt 0000 lanosterol 119901 = 0011)

4 Discussion

41 ApoE4 Mice as Model for the Early Asymptomatic Phasein AD In this study we investigated the extent to whichapoE4 mice display cerebrovascular flaws synaptic loss andconnectivity during aging The 1205764 allele of the apoE gene isstrongly associated with sporadic AD [45] Among severalproposed mechanisms by which apoE4 promotes AD thereare indications that apoE4 is less effective in synaptic repairand remodelling processes compared to other isoforms [4647] Moreover apoE4 carriers are clearly more susceptibleto vascular brain damage (eg stroke brain haemorrhage[29 30 48]) and they display aberrant functional connectiv-ity [49]

Similarly the apoE4 mouse line exhibits increased riskof developing vascular disorders and neuronal deficits dueto altered cholesterol metabolism especially when chal-lenged with a high-fat diet [33] Recently we also describedspontaneous functional connectivity deficits in these micepossibly associated with cerebral blood flow reduction [50]

Because these deficits seem to be aggravated with aging weinvestigated this mouse line at two different ages

At 10ndash12 months of age apoE4mice did not display manydifferences compared to WT animals Despite some slightalterations in the sterol levels of 12-month-old apoE4 miceall other measured parameters including cerebral blood flowand number of postsynapses were unaffected compared toWT animals Previously we have shown that at this age alsocerebral blood volume amount of presynaptic boutons andneurogenesis did not differ from WT mice [35] The lack ofcerebrovascular alterations like a reduced CBF at this agemayexplain the absence of pathologies

However at 16ndash18 months of age apoE4 mice revealedreduced CBF and accelerated neurodegeneration which aretypical features of prodromal AD [51ndash54] Specifically wedetected reduced thalamic and cortical perfusion reducedcortical postsynaptic density increased cortical mean dif-fusivity (MD) and reduced fractional anisotropy (FA) inwhite matter tracts Similar changes in brain diffusivity asa biomarker for white and gray matter integrity have beenreported in human apoE4 carriers [55ndash58] These structuralmodifications may be linked to the isoform-specific roleof apoE in synaptic development dendrite formation andaxonal guidance which to some extent may be impairedin apoE4 carriers [59] Nevertheless these results were notconsistent across different ages suggesting that changes inWMGM microstructure properties may not directly reflectan associated AD-like pathology [60] Moreover at this agewe measured a widespread reduction in functional connec-tivity at rest which was previously reported [17]

Overall in line with other studies in this mouse model[34] these findings suggest that the apoE4 mice sponta-neously develop age- and apoE4-dependent accelerated brainpathologyThis is in agreement with human studies on apoE4carriers showing a faster decline of CBF during aging [61 62]and reduced connectivity between cortical regions at old age

Neural Plasticity 11

[63ndash65] However the apoE4 mice exhibited relatively smallgenotype effects like CBF decline and reduced connectivityjust at 18months of age and not at 12months of ageThereforeone could argue that an 18-month-old apoE4 mouse is atmost similar to a 65-year-old still healthy possibly onlymildlyimpaired human apoE4 carrier showing no signs of dementiayet and carrying no multiple risk factors and comorbiditiesIt has been found namely that human apoE4 carriers oftencarry multiple risk factors (genetic modifiers comorbiditiesand lifestyle factors) that contribute significantly to synapticintegrity The only risk factor included in our mice was agecombined with apoE4 genotype causing mild pathology

A recent review on apoE-related biomarker profiles inthe early phase of AD further elucidates this relatively novelconcept of the apoE4 to be consideredmore as a vulnerabilityfactor rather than a pathogenic factor [66] Based on thisidea it is the interaction between these vulnerabilities andthe age-related pathological events that may trigger synapticloss contributing to the development of AD In our studythis aging-dependency of apoE4 seemed confirmed as mostof the biomarkers for brain deficits were identified onlyin the 18-month-old apoE4 group representing an earlystage of the disease when complete early AD-like pathologyis still not fully developed Holding this hypothesis themodel becomes extremely attractive to study the effects ofnutritional intervention as a preventive strategy against earlyAD-like pathology

42 Dietary Intervention In the current experiment wefed the mice a specific multinutrient supplementation dietdesigned to ameliorate synapse loss and to reducemembrane-related pathology in AD by providing nutritional precursorsand cofactors to support neuronal membrane formation andfunction [67] This nutrient combination called Fortasyncomprises uridine docosahexaenoic acid eicosapentaenoicacid choline phospholipids folic acid vitamins B12 B6C and E and selenium Some of the components in thesediets such as omega-3 long-chain polyunsaturated fatty acids(n3- LCPUFAs) have also been shown to improve vascularhealth [5 68ndash71]The results confirmed our initial hypothesisthat the Fortasyn diet has the potential to reduce the occur-rence of vulnerabilities for AD by simultaneously improv-ing cerebrovascular health and enhancing neuroprotectivemechanisms However it is also possible that the capacity ofthe diet to support membrane phospholipid synthesis couldunderlie both the synapse formationfunctional connectivityand the effects on cerebrovascular functioning Besides thecombination of phosphatide precursors like n3-LCPUFAsuridine and choline has proven to synergistically increase thesynthesis of synaptic proteins and phospholipids in the brain[14ndash16]

These findings confirm novel important mechanisms bywhich these diets may affect AD onset and developmentsimilar to earlier findings in a transgenic AD mouse model[17] Particularly among the different parameters analysedthe strongest and most consistent dietary effect in these stud-ies involves the improvement of cerebrovascular health andfunctional connectivity Several epidemiological studies andcontrolled trials showed a correlation between B vitamins

n3-LCPUFAs and MUFA (like oleic acid from olive oiland nuts) and improvements in autonomic function low-ered blood pressure reduced atherosclerosis reduced totalhomocysteine and enhanced microvascular endothelium-dependent vasodilation processes [4 72 73]

All these factors may contribute to an improved func-tionality of the brain vasculature with a beneficial effectparticularly in apoE4 mice in which these pathologies areaggravated Interestingly increased cortical CBF and levelsof postsynaptic markers were found in both apoE4 andWT animals on Fortasyn diet these findings suggest thatthe diet had a similar effect in both genotypes and itscontribution is not determined by the apoE genotype Wehave also shown that the Fortasyn diet affected brain fattyacid profiles in both genotypes (Supplementary MaterialTable 1) by decreasing the relative concentration of n6-fatty acids (notably arachidonic acid) which is significantlyincreased in the 18-month-old apoE4 mice compared toWT and by increasing the relative concentration of n3-fatty acids (notably DHA) and monounsaturated fatty acidsespecially oleic acid These supplementary results indicatea replacement of n6 fatty acids from cell membranes infavour of n3-LCPUFAs (reflected by the increased n3n6ratio) which has beneficial effects on membrane fluidity andneuronal transmission and signalling [74ndash76] Furthermorewe have shown that the Fortasyn diet similarly affectedserum (Supplementary Material Table 2) and brain sterollevel (Supplementary Material Table 3) In detail in theplasma of aged animals on Fortasyn we found decreasedlevels of cholesterol but also of precursors like lanosterol andderivates of cholesterol like 24OH-cholesterol Notably inthe cerebellum of aged animals on Fortasyn we detected adecreased concentration of the cholesterol precursor lanos-terol and increased level of a derivate of cholesterol 24S-hydroxycholesterol In the brain the enzyme cholesterol 24S-hydroxylase converts cholesterol to 24S-hydroxycholesterolThis mechanism is the most important pathway for theelimination of brain cholesterol and the maintenance ofbrain cholesterol homeostasis [77ndash80] These results arealso in line with our previous study using another ADmouse model overexpressing A120573 Here another cholesterolprecursor lathosterol was decreased in the brain of Fortasynfed AD and WT mice while again an increased cerebrallevel of the derivate of cholesterol 24S-hydroxycholesterolwas measured in Fortasyn fed AD and WT mice [81] Ourdata indicate a higher elimination rate of brain cholesterol inFortasyn fed mice

Thismay also explain the improvedwhitematter integrityand preserved functional connectivity in both WT and ADmice fed with Fortasyn In support of this hypothesis it hasbeen demonstrated that animals fed with a diet containinguridine monophosphate (UMP) n3-LCPUFAs and cholineshowed increased levels of brain phospholipids dendriticspines and neurite outgrowth [14ndash16]

5 Conclusions

Overall the study presented here further proved thattwo simultaneous protective mechanisms on vascular and


synapse health are both enhanced by the specific Fortasyndiet and may strengthen each other synergistically indepen-dent of the apoE genotype The beneficial effect of thesediets is suggested to be caused by increased productionof phospholipids to sustain synaptic genesis and repairprocesses [14 82] In this and other recent experiments weshowed that a strong effect of these diets also involves theamelioration of cerebrovascular health Although decreasedcerebral perfusion has been recognized as an early andimportant contributor to ADpathology and cognitive decline[83] we believe that this aspect is not sufficiently consideredin human nutritional intervention studies In our mice wehave detected most of the pathological effects of apoE4 justat 18 months of age and therefore most of the beneficialeffects of the diet were just present at 18 months of ageIt is important to stress that our apoE4 mouse model onlyrepresents susceptibility to cognitive impairment and just likein human apoE4 carriers multiple risk factors are required incombination with apoE4 to precipitate disease pathology Forfuture research it would therefore be interesting to study theeffect of the Fortasyn diet in older (24 months of age) apoE4mice in apoE4 mice with induced apoE4 comorbidities likehypertension stroke or in apoE4 mice on high-fat diet orin female apoE4 mice resembling more closely the humansusceptibility to AD For example in clinical studies on brainatrophy and clinical decline among cognitively normal olderindividuals and individuals with mild cognitive impairmentand Alzheimerrsquos disease it has been shown that the presenceof apoE4 significantly accelerated rates of decline andwomenin all cohorts had higher rates of decline than men [84]Additionally a preclinical study revealed that expression ofhuman apoE4 renders aged mice fed with a western-typediet more susceptible to sensorimotor deficits upon strokeindicating an altered functional outcome following stroke inapoE4 carriers [85] Furthermore this study demonstratedthe value of a multimodal approach including advanced MRneuroimaging tools for detecting changes in brain structureand function with respect to dietary intervention

Abbreviations

AD Alzheimerrsquos diseaseCBF Cerebral blood flowFC Functional connectivityWT Wild-typen3-LCPUFAs Omega-3 long-chain polyunsaturated

fatty acidsDHA Docosahexaenoic acidEPA Eicosapentaenoic acidapoE ApolipoproteinA120573 Amyloid-120573ES Embryonic stemGE Gradient echoMD Mean water diffusivityFA Fractional anisotropyPBS Phosphate buffered salineIML Inner molecular layerOML Outer molecular layer

CA Cornus ammonisGC-MS-SIM Gas-chromatography-mass-

spectrometry-selected-ion-monitoring

Competing Interests

LausM Broersen is employed by Nutricia AdvancedMedicalNutrition Danone Research The rest of the authors declarethat there are no actual or potential competing interests

Authorsrsquo Contributions

Valerio Zerbi andMaximilianWiesmann contributed equallyto the present work and share first authorship

Acknowledgments

This work was supported by the European CommunityrsquosSeventh Framework Programme (EU 7th Framework Lipi-diDiet project) (FP72007ndash2013) under Grant Agreementno 211696 and NWO Investment Grants 91106021 and BIG(VISTA)This study was supported by a grant (no11528) fromthe Internationale Stichting Alzheimer Onderzoek (ISAO)The authors thank Tim Emmerzaal Jos Dederen MaartjeMutsaers Maarten van Beek Karin Vos Sabine DenissenRobert van der Lugt Ilse Arnoldussen and Laura Mellendijkfor their laboratory work and Bianca Lemmers-van deWeemKitty Lemmens-Hermans Iris Lamers-Elemans and HenkArnts for their excellent care of their mice

References

[1] F Sofi F Cesari R Abbate G F Gensini and A CasinildquoAdherence to Mediterranean diet and health status meta-analysisrdquo British Medical Journal vol 337 p a1344 2008

[2] N Scarmeas Y Stern M-X Tang R Mayeux and J ALuchsinger ldquoMediterranean diet and risk for Alzheimerrsquos dis-easerdquo Annals of Neurology vol 59 no 6 pp 912ndash921 2006

[3] N Scarmeas J A Luchsinger R Mayeux and Y SternldquoMediterranean diet and Alzheimer disease mortalityrdquo Neurol-ogy vol 69 no 11 pp 1084ndash1093 2007

[4] R Estruch E Ros J Salas-Salvado et al ldquoPrimary preventionof cardiovascular disease with a Mediterranean dietrdquo The NewEngland Journal of Medicine vol 368 no 14 pp 1279ndash12902013

[5] J H Lee J H OrsquoKeefe C J Lavie R Marchioli and W SHarris ldquoOmega-3 fatty acids for cardioprotectionrdquoMayo ClinicProceedings vol 83 no 3 pp 324ndash332 2008

[6] C Iadecola ldquoNeurovascular regulation in the normal brain andin Alzheimerrsquos diseaserdquoNature Reviews Neuroscience vol 5 no5 pp 347ndash360 2004

[7] F Thies J M C Garry P Yaqoob et al ldquoAssociation of n-3 polyunsaturated fatty acids with stability of atheroscleroticplaques a randomised controlled trialrdquoThe Lancet vol 361 no9356 pp 477ndash485 2003

[8] J M Geleijnse E J Giltay D E Grobbee A R T Donders andF J Kok ldquoBlood pressure response to fish oil supplementationmetaregression analysis of randomized trialsrdquo Journal of Hyper-tension vol 20 no 8 pp 1493ndash1499 2002


[9] M LDaviglus J Stamler A J Orencia et al ldquoFish consumptionand the 30-year risk of fatal myocardial infarctionrdquo The NewEngland Journal ofMedicine vol 336 no 15 pp 1046ndash1053 1997

[10] F B Hu L BronnerW CWillett et al ldquoFish and omega-3 fattyacid intake and risk of coronary heart disease in womenrdquo TheJournal of the American Medical Association vol 287 no 14 pp1815ndash1821 2002

[11] C von Schacky and W S Harris ldquoCardiovascular benefits ofomega-3 fatty acidsrdquo Cardiovascular Research vol 73 no 2 pp310ndash315 2007

[12] R J Wurtman M Cansev T Sakamoto and I Ulus ldquoNutri-tional modifiers of aging brain function use of uridine andother phosphatide precursors to increase formation of brainsynapsesrdquo Nutrition Reviews vol 68 no 2 pp S88ndashS101 2010

[13] S B Weiss and E P Kennedy ldquoThe function of cytidinecoenzymes in the biosynthesis of phospholipidesrdquo The Journalof Biological Chemistry vol 222 no 1 pp 193ndash214 1956

[14] R J Wurtman I H Ulus M Cansev C J Watkins LWang and G Marzloff ldquoSynaptic proteins and phospholipidsare increased in gerbil brain by administering uridine plusdocosahexaenoic acid orallyrdquoBrain Research vol 1088 no 1 pp83ndash92 2006

[15] M Cansev and R J Wurtman ldquoChronic administration ofdocosahexaenoic acid or eicosapentaenoic acid but not arachi-donic acid alone or in combination with uridine increasesbrain phosphatide and synaptic protein levels in gerbilsrdquo Neu-roscience vol 148 no 2 pp 421ndash431 2007

[16] T Sakamoto M Cansev and R J Wurtman ldquoOral sup-plementation with docosahexaenoic acid and uridine-51015840-monophosphate increases dendritic spine density in adult gerbilhippocampusrdquo Brain Research vol 1182 no 1 pp 50ndash59 2007

[17] V Zerbi D Jansen M Wiesmann et al ldquoMultinutrient dietsimprove cerebral perfusion and neuroprotection in a murinemodel of Alzheimerrsquos diseaserdquo Neurobiology of Aging vol 35no 3 pp 600ndash613 2014

[18] M Wiesmann D Jansen V Zerbi L M Broersen A Gartheand A J Kiliaan ldquoImproved spatial learning strategy andmemory in aged Alzheimer AbetaPPswePS1dE9 mice on amulti-nutrient dietrdquo Journal of Alzheimerrsquos Disease vol 37 no1 pp 233ndash245 2013

[19] P J G H Kamphuis and P Scheltens ldquoCan nutrients preventor delay onset of Alzheimerrsquos diseaserdquo Journal of AlzheimerrsquosDisease vol 20 no 3 pp 765ndash775 2010

[20] J L Cummings ldquoFood for thought Souvenaid in mildAlzheimerrsquos diseaserdquo Journal of Alzheimerrsquos Disease vol 31 no1 pp 237ndash238 2012

[21] P Scheltens P J G H Kamphuis F R J Verhey et al ldquoEfficacyof a medical food in mild Alzheimerrsquos disease a randomizedcontrolled trialrdquo Alzheimerrsquos amp Dementia vol 6 no 1 pp 1e1ndash10e1 2010

[22] P Scheltens J W R Twisk R Blesa et al ldquoEfficacy of souve-naid in mild alzheimerrsquos disease results from a randomizedcontrolled trialrdquo Journal of Alzheimerrsquos Disease vol 31 no 1 pp225ndash236 2012

[23] H de Waal C J Stam M M Lansbergen et al ldquoThe effect ofsouvenaid on functional brain network organisation in patientswith mild Alzheimerrsquos disease a randomised controlled studyrdquoPLoS ONE vol 9 no 1 Article ID e86558 2014

[24] A M Minihane S Khan E C Leigh-Firbank et al ldquoApoEpolymorphism and fish oil supplementation in subjects with anatherogenic lipoprotein phenotyperdquo Arteriosclerosis Thrombo-sis and Vascular Biology vol 20 no 8 pp 1990ndash1997 2000

[25] L J Whalley I J Deary J M Starr et al ldquon-3 Fatty acid ery-throcyte membrane content APOE 1205764 and cognitive variationan observational follow-up study in late adulthoodrdquo AmericanJournal of Clinical Nutrition vol 87 no 2 pp 449ndash454 2008

[26] C Samieri S Lorrain B Buaud et al ldquoRelationship betweendiet and plasma long-chain n-3 PUFAs in older people impactof apolipoprotein E genotyperdquo Journal of Lipid Research vol 54no 9 pp 2559ndash2567 2013

[27] V I Zannis J L BreslowGUtermann et al ldquoProposed nomen-clature of apoE isoproteins apoE genotypes and phenotypesrdquoJournal of Lipid Research vol 23 no 6 pp 911ndash914 1982

[28] C Frieden and K Garai ldquoStructural differences between apoE3and apoE4 may be useful in developing therapeutic agents forAlzheimerrsquos diseaserdquo Proceedings of the National Academy ofSciences of theUnited States of America vol 109 no 23 pp 8913ndash8918 2012

[29] C C Liu T Kanekiyo H Xu and G Bu ldquoApolipoprotein Eand Alzheimer disease risk mechanisms and therapyrdquo NatureReviews Neurology vol 9 no 2 pp 106ndash118 2013

[30] B V Zlokovic ldquoCerebrovascular effects of apolipoprotein Eimplications for Alzheimer diseaserdquo JAMA Neurology vol 70no 4 pp 440ndash444 2013

[31] K Rannikmae R N Kalaria S M Greenberg et alldquoAPOE associations with severe CAA-associated vasculopathicchanges collaborative meta-analysisrdquo Journal of NeurologyNeurosurgery and Psychiatry vol 85 no 3 pp 300ndash305 2014

[32] K Hultman S Strickland and E H Norris ldquoThe APOE12057641205764 genotype potentiates vascular fibrin(ogen) depositionin amyloid-laden vessels in the brains of Alzheimerrsquos diseasepatientsrdquo Journal of Cerebral Blood Flow and Metabolism vol33 no 8 pp 1251ndash1258 2013

[33] C Knouff M E Hinsdale H Mezdour et al ldquoApo E structuredetermines VLDL clearance and atherosclerosis risk in micerdquoJournal of Clinical Investigation vol 103 no 11 pp 1579ndash15861999

[34] J Yin G H Turner S W Coons M Maalouf E M Reimanand J Shi ldquoAssociation of amyloid burden brain atrophy andmemory deficits in aged apolipoprotein epsilon4micerdquoCurrentAlzheimer Research vol 11 no 3 pp 283ndash290 2014

[35] D Jansen V Zerbi C I Janssen et al ldquoImpact of a multi-nutrient diet on cognition brain metabolism hemodynamicsand plasticity in apoE4 carrier and apoE knockout micerdquo BrainStructure and Function vol 219 no 5 pp 1841ndash1868 2014

[36] P M Sullivan H Mezdour Y Aratani et al ldquoTargetedreplacement of the mouse apolipoprotein E gene with thecommon human APOE3 allele enhances diet-induced hyper-cholesterolemia and atherosclerosisrdquo The Journal of BiologicalChemistry vol 272 no 29 pp 17972ndash17980 1997

[37] P G Reeves F H Nielsen and G C Fahey Jr ldquoAIN-93 purifieddiets for laboratory rodents final report of the American Insti-tute ofNutrition adhocwriting committee on the reformulationof the AIN-76A rodent dietrdquo The Journal of Nutrition vol 123no 11 pp 1939ndash1951 1993

[38] S-G Kim ldquoQuantification of relative cerebral blood flowchange by flow-sensitive alternating inversion recovery (FAIR)technique application to functional mappingrdquo Magnetic Reso-nance in Medicine vol 34 no 3 pp 293ndash301 1995

[39] L-A Harsan D Paul S Schnell et al ldquoIn vivo diffusion tensormagnetic resonance imaging and fiber tracking of the mousebrainrdquo NMR in Biomedicine vol 23 no 7 pp 884ndash896 2010


[40] M P Zwiers ldquoPatching cardiac and head motion artefacts indiffusion-weighted imagesrdquoNeuroImage vol 53 no 2 pp 565ndash575 2010

[41] G Paxinos and K Franklin The Mouse Brain in StereotaxicCoordinates Academic Press San Diego Calif USA 2001

[42] S J Sawiak N I Wood G B Williams A J Morton and T ACarpenter ldquoVoxel-based morphometry in the R62 transgenicmouse reveals differences between genotypes not seen withmanual 2D morphometryrdquo Neurobiology of Disease vol 33 no1 pp 20ndash27 2009

[43] E Jonckers J vanAudekerke G deVisscher A van der Lindenand M Verhoye ldquoFunctional connectivity fMRI of the rodentbrain comparison of functional connectivity networks in ratand mouserdquo PLoS ONE vol 6 no 4 Article ID e18876 2011

[44] D Lutjohann A Brzezinka E Barth et al ldquoProfile ofcholesterol-related sterols in aged amyloid precursor proteintransgenic mouse brainrdquo Journal of Lipid Research vol 43 no7 pp 1078ndash1085 2002

[45] R W Mahley and S C Rall Jr ldquoApolipoprotein E far morethan a lipid transport proteinrdquo Annual Review of Genomics andHuman Genetics vol 1 pp 507ndash537 2000

[46] RWMahley K HWeisgraber and Y Huang ldquoApolipoproteinE4 a causative factor and therapeutic target in neuropathologyincluding Alzheimerrsquos diseaserdquo Proceedings of the NationalAcademy of Sciences of the United States of America vol 103 no15 pp 5644ndash5651 2006

[47] P B Verghese J M Castellano and D M HoltzmanldquoApolipoprotein E inAlzheimerrsquos disease and other neurologicaldisordersrdquoTheLancet Neurology vol 10 no 3 pp 241ndash252 2011

[48] B V Zlokovic ldquoNeurovascular pathways to neurodegenerationin Alzheimerrsquos disease and other disordersrdquo Nature ReviewsNeuroscience vol 12 no 12 pp 723ndash738 2011

[49] Y I Sheline J C Morris A Z Snyder et al ldquoAPOE4 allele dis-rupts resting state fMRI connectivity in the absence of amyloidplaques or decreased CSF A12057342rdquo Journal of Neuroscience vol30 no 50 pp 17035ndash17040 2010

[50] V Zerbi M Wiesmann T L Emmerzaal et al ldquoResting-statefunctional connectivity changes in aging apoE4 and apoE-KOmicerdquo The Journal of Neuroscience vol 34 no 42 pp 13963ndash13975 2014

[51] Z Wang S R Das S X Xie S E Arnold J A Detre and DA Wolk ldquoArterial spin labeled MRI in prodromal Alzheimerrsquosdisease a multi-site studyrdquo NeuroImage Clinical vol 2 no 1pp 630ndash636 2013

[52] P Vemuri H JWiste S DWeigand et al ldquoSerial MRI and CSFbiomarkers in normal aging MCI and ADrdquo Neurology vol 75no 2 pp 143ndash151 2010

[53] C R Jack Jr D S KnopmanW J Jagust et al ldquoTracking patho-physiological processes in Alzheimerrsquos disease an updatedhypothetical model of dynamic biomarkersrdquo The Lancet Neu-rology vol 12 no 2 pp 207ndash216 2013

[54] C R Jack Jr V J Lowe SDWeigand et al ldquoSerial PIB andMRIin normal mild cognitive impairment and Alzheimers diseaseimplications for sequence of pathological events in Alzheimersdiseaserdquo Brain vol 132 no 5 pp 1355ndash1365 2009

[55] V Heise N Filippini A J Trachtenberg S Suri K P Ebmeierand C E Mackay ldquoApolipoprotein E genotype gender and agemodulate connectivity of the hippocampus in healthy adultsrdquoNeuroImage vol 98 pp 23ndash30 2014

[56] V Heise N Filippini K P Ebmeier and C E MacKay ldquoTheAPOE 1205764 allele modulates brain white matter integrity in

healthy adultsrdquoMolecular Psychiatry vol 16 no 9 pp 908ndash9162011

[57] J Nierenberg N Pomara M J Hoptman J J Sidtis B AArdekani and K O Lim ldquoAbnormal white matter integrity inhealthy apolipoprotein E epsilon4 carriersrdquo NeuroReport vol16 no 12 pp 1369ndash1372 2005

[58] R A Honea E Vidoni A Harsha and J M Burns ldquoImpact ofAPOE on the healthy aging brain a voxel-based MRI and DTIstudyrdquo Journal of Alzheimerrsquos Disease vol 18 no 3 pp 553ndash5642009

[59] J Kim J M Basak and D M Holtzman ldquoThe role ofapolipoprotein E in Alzheimerrsquos diseaserdquo Neuron vol 63 no3 pp 287ndash303 2009

[60] L TWestlye I Reinvang H Rootwelt and T Espeseth ldquoEffectsof APOE on brain white matter microstructure in healthyadultsrdquo Neurology vol 79 no 19 pp 1961ndash1969 2012

[61] N Scarmeas andY Stern ldquoImaging studies andAPOE genotypein persons at risk for Alzheimerrsquos diseaserdquo Current PsychiatryReports vol 8 no 1 pp 11ndash17 2006

[62] C E Wierenga L R Clark S I Dev et al ldquoInteraction of ageand APOE genotype on cerebral blood flow at restrdquo Journal ofAlzheimerrsquos Disease vol 34 no 4 pp 921ndash935 2013

[63] J L OrsquoBrien K M OrsquoKeefe P S Laviolette et al ldquoLongitudinalfMRI in elderly reveals loss of hippocampal activation withclinical declinerdquo Neurology vol 74 no 24 pp 1969ndash1976 2010

[64] M M Machulda D T Jones P Vemuri et al ldquoEffect of APOEepsilon4 status on intrinsic network connectivity in cognitivelynormal elderly subjectsrdquoArchives of Neurology vol 68 no 9 pp1131ndash1136 2011

[65] J A Brown K H Terashima A C Burggren et al ldquoBrainnetwork local interconnectivity loss in aging APOE-4 allelecarriersrdquo Proceedings of the National Academy of Sciences of theUnited States of America vol 108 no 51 pp 20760ndash20765 2011

[66] I Reinvang T Espeseth and L T Westlye ldquoAPOE-relatedbiomarker profiles in non-pathological aging and early phasesof Alzheimerrsquos diseaserdquo Neuroscience and BiobehavioralReviews vol 37 no 8 pp 1322ndash1335 2013

[67] N van Wijk L M Broersen M C De Wilde et al ldquoTargetingsynaptic dysfunction in Alzheimerrsquos disease by administering aspecific nutrient combinationrdquo Journal of Alzheimerrsquos Diseasevol 38 no 3 pp 459ndash479 2014

[68] C Dawczynski L Martin A Wagner and G Jahreis ldquon-3 LC-PUFA-enriched dairy products are able to reduce cardiovas-cular risk factors a double-blind cross-over studyrdquo ClinicalNutrition vol 29 no 5 pp 592ndash599 2010

[69] E I Lev A Solodky N Harel et al ldquoTreatment of aspirin-resistant patients with omega-3 fatty acids versus aspirin doseescalationrdquo Journal of the American College of Cardiology vol55 no 2 pp 114ndash121 2010

[70] V Schiano E Laurenzano G Brevetti et al ldquoOmega-3 polyun-saturated fatty acid in peripheral arterial disease effect on lipidpattern disease severity inflammation profile and endothelialfunctionrdquo Clinical Nutrition vol 27 no 2 pp 241ndash247 2008

[71] A Zampelas ldquoEicosapentaenoic acid (EPA) from highly con-centrated n-3 fatty acid ethyl esters is incorporated intoadvanced atherosclerotic plaques and higher plaque EPA isassociated with decreased plaque inflammation and increasedstabilityrdquo Atherosclerosis vol 212 no 1 pp 34ndash35 2010

[72] A D Dangour P J Whitehouse K Rafferty et al ldquoB-vitamins and fatty acids in the prevention and treatment ofAlzheimerrsquos disease and dementia a systematic reviewrdquo Journalof Alzheimerrsquos Disease vol 22 no 1 pp 205ndash224 2010


[73] S M Artham C J Lavie R V Milani R G Anand J HOrsquoKeefe and H O Ventura ldquoFish oil in primary and secondarycardiovascular preventionrdquo Ochsner Journal vol 8 no 2 pp49ndash60 2008

[74] J-M Bourre M Francois A Youyou et al ldquoThe effects ofdietary 120572-linolenic acid on the composition of nerve mem-branes enzymatic activity amplitude of electrophysiologicalparameters resistance to poisons and performance of learningtasks in ratsrdquo Journal of Nutrition vol 119 no 12 pp 1880ndash18921989

[75] J M Bourre O Dumont M Piciotti et al ldquoEssentiality ofomega 3 fatty acids for brain structure and functionrdquo WorldReview of Nutrition and Dietetics vol 66 pp 103ndash117 1991

[76] E Farkas M C De Wilde A J Kiliaan J Meijer J N Keijserand P G M Luiten ldquoDietary long chain PUFAs differentiallyaffect hippocampal muscarinic 1 and serotonergic 1A receptorsin experimental cerebral hypoperfusionrdquo Brain Research vol954 no 1 pp 32ndash41 2002

[77] D Lutjohann O Breuer G Ahlborg et al ldquoCholesterol home-ostasis in human brain evidence for an age-dependent fluxof 24S-hydroxycholesterol from the brain into the circulationrdquoProceedings of the National Academy of Sciences of the UnitedStates of America vol 93 no 18 pp 9799ndash9804 1996

[78] E G Lund J M Guileyardo and D W Russell ldquocDNAcloning of cholesterol 24-hydroxylase a mediator of cholesterolhomeostasis in the brainrdquo Proceedings of the National Academyof Sciences of the United States of America vol 96 no 13 pp7238ndash7243 1999

[79] J Koschack D Lutjohann C Schmidt-Samoa and E IrleldquoSerum 24S-hydroxycholesterol and hippocampal size inmiddle-agednormal individualsrdquoNeurobiology of Aging vol 30no 6 pp 898ndash902 2009

[80] I Bjorkhem D Lutjohann O Breuer A Sakinis and AWennmalm ldquoImportance of a novel oxidative mechanism forelimination of brain cholesterol Turnover of cholesterol and24(S)-hydroxycholesterol in rat brain as measured with 18O2techniques in vivo and in vitrordquo Journal of Biological Chemistryvol 272 no 48 pp 30178ndash30184 1997

[81] D Jansen V Zerbi I A C Arnoldussen et al ldquoEffects ofspecific multi-nutrient enriched diets on cerebral metabolismcognition and neuropathology in A120573PPswe-PS1dE9 micerdquoPLoS ONE vol 8 no 9 Article ID e75393 2013

[82] R J Wurtman M Cansev T Sakamoto and I H Ulus ldquoUseof phosphatide precursors to promote synaptogenesisrdquo AnnualReview of Nutrition vol 29 pp 59ndash87 2009

[83] R Altman and J C Rutledge ldquoThe vascular contribution toAlzheimerrsquos diseaserdquo Clinical Science vol 119 no 10 pp 407ndash421 2010

[84] D Holland R S Desikan A M Dale and L K McEvoyldquoHigher rates of decline for women and apolipoprotein E 1205764carriersrdquoAmerican Journal of Neuroradiology vol 34 no 12 pp2287ndash2293 2013

[85] H Dhungana T Rolova E Savchenko et al ldquoWestern-typediet modulates inflammatory responses and impairs functionaloutcome following permanent middle cerebral artery occlusionin aged mice expressing the human apolipoprotein E4 allelerdquoJournal of Neuroinflammation vol 10 article 102 2013

Submit your manuscripts athttpwwwhindawicom

Neurology Research International

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Alzheimerrsquos DiseaseHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

International Journal of

ScientificaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014


BioMed Research International


Research and TreatmentSchizophrenia

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014


Neural Plasticity


Parkinsonrsquos Disease


Research and TreatmentAutism

Sleep DisordersHindawi Publishing Corporationhttpwwwhindawicom Volume 2014


Neuroscience Journal

Epilepsy Research and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014


Psychiatry Journal


Computational and Mathematical Methods in Medicine

Depression Research and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014


Brain ScienceInternational Journal of

StrokeResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Neurodegenerative Diseases


Journal of

Cardiovascular Psychiatry and NeurologyHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Research ArticleA Dietary Treatment Improves Cerebral Blood Flow andBrain Connectivity in Aging apoE4 Mice

Maximilian Wiesmann12 Valerio Zerbi13 Diane Jansen1 Roy Haast1 Dieter Luumltjohann4

Laus M Broersen56 Arend Heerschap3 and Amanda J Kiliaan1

1Department of Anatomy Donders Institute for Brain Cognition amp Behaviour Radboud University Medical Center6525 EZ Nijmegen Netherlands2Department of Geriatric Medicine Radboud University Medical Center 6525 EZ Nijmegen Netherlands3Department of Radiology and Nuclear Medicine Radboud University Medical Center 6525 EZ Nijmegen Netherlands4Institute for Clinical Chemistry and Clinical Pharmacology University of Bonn Bonn Germany5Nutricia Research Advanced Medical Nutrition Utrecht Netherlands6UIPS Utrecht University Utrecht Netherlands

Correspondence should be addressed to Amanda J Kiliaan amandakiliaanradboudumcnl

Received 8 January 2016 Accepted 22 February 2016

Academic Editor Tae-Jin Kim

Copyright copy 2016 Maximilian Wiesmann et al This is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in any medium provided the original work is properlycited

APOE 1205764 (apoE4) polymorphism is the main genetic determinant of sporadic Alzheimerrsquos disease (AD) A dietary approach(Fortasyn) including docosahexaenoic acid eicosapentaenoic acid uridine choline phospholipids folic acid vitamins B12 B6C and E and selenium has been proposed for dietary management of AD We hypothesize that the diet could inhibit AD-likepathologies in apoE4 mice specifically cerebrovascular and connectivity impairment Moreover we evaluated the diet effect oncerebral blood flow (CBF) functional connectivity (FC) graywhite matter integrity and postsynaptic density in aging apoE4miceAt 10ndash12 months apoE4 mice did not display prominent pathological differences compared to wild-type (WT) mice However16ndash18-month-old apoE4 mice revealed reduced CBF and accelerated synaptic loss The diet increased cortical CBF and amountof synapses and improved white matter integrity and FC in both aging apoE4 and WT mice We demonstrated that protectivemechanisms on vascular and synapse health are enhanced by Fortasyn independent of apoE genotype We further showed theefficacy of a multimodal translational approach including advanced MR neuroimaging to study dietary intervention on brainstructure and function in aging

1 Introduction

Extensive research has been pursued in search of an effectivetherapy for Alzheimerrsquos disease (AD) However no treatmentis yet available nor does it seem near Preventive approacheshave therefore consistently emerged as key policy prioritiesin recently formulated dementia strategiesThese approachesinclude modification of health-compromising behavior suchas lifestyle and dietary intake that may lead to AD Forexample the Mediterranean diet (high consumption of fruitvegetables and legumes moderate consumption of fish nutsand olive oil as the main source of fats) has been associatedwith a reduced risk of AD [1 2] and with a lower mortality

[3 4] The mechanisms via which diet influences the onsetand progression ofADpathology are still under investigationOne possible mode of action is the beneficial effect ofnutrients such as omega-3 long-chain polyunsaturated fattyacids (n3-LCPUFAs) on the vascular system [4 5]

Herewith n3-LCPUFAs will target the very early asymp-tomatic phase of the disease in which (cerebro)vascularimpairment is the strongest contributor to the onset andprogression of neurodegenerative traits of typical AD anddementia in general [6] n3-LCPUFAs may diminish severityof vascular risk factors like atherosclerosis [7] high bloodpressure [8] and other cardiovascular diseases [7 9ndash11]which are risk factors for AD as well Other nutrients may

Hindawi Publishing CorporationNeural PlasticityVolume 2016 Article ID 6846721 15 pageshttpdxdoiorg10115520166846721

2 Neural Plasticity










Neural Plasticity 3



Control[g100 g]

Fortasyn[g100 g]


2


BirthStart diets

0 2 4 10 12 14 16 18


Months






2








4 Neural Plasticity








3 Results


31 Body Weight




Neural Plasticity 5

ControlFortasyn

lowast

lowastlowast


lowastlowastlowast



0

10

20

30

40

50

60

70Bo

dy w

eigh

t (g)

(a)

ControlFortasyn

lowast

lowastlowast

lowast

lowastlowast



0

10

20

30

40

50

60

70

Body

wei

ght (

g)

(b)












323 rsfMRI




6 Neural Plasticity

12-month-oldlowast

0

100

200

300



Wild-type

Wild-type

CBF

(mL100

gm

in)

ControlFortasyn

(a)

0

100

200

300


18-month-old

lowast lowast

lowast


Wild-type

Wild-type

CBF

(mL100

gm

in)

ControlFortasyn

lowast

(b)












MD


12-month-old

ApoE

4W

ild-ty

pe

tractOptic

Wild

-type

ApoE

4

CCminus07mm

Wild

-type

ApoE

4

CCminus17mm

ApoE

4W

ild-ty

pe

CCminus22mm

ApoE

4W

ild-ty

pe

Fornix

02

04

06

08

Frac

tiona

l ani

sotro

py (F

A)

lowastlowast

ControlFortasyn

lowast

lowastlowast

00006

00007

00008

00009

00010

Mea

n di

ffusiv

ity (M

D) (

mm

2s

)

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

Motorcortex

Auditorycortex

Sensorycortex

Visualcortex

ApoE

4W

ild-ty

pe

Hippocampus

(b)

ControlFortasyn

lowastlowast

00006

00007

00008

00009

00010

Mea

n di

ffusiv

ity (M

D) (

mm

2s

)

Wild

-type

ApoE

4

Motorcortex

Wild

-type

ApoE

4

Auditorycortex

Wild

-type

ApoE

4

Sensorycortex

Wild

-type

ApoE

4Visualcortex

Wild

-type

ApoE

4

Hippocampus

18-month-old

lowastlowast

ApoE

4W

ild-ty

pe

Fornix

Wild

-type

ApoE

4

CCminus22mm

Wild

-type

ApoE

4

CCminus17mm

Wild

-type

ApoE

4

CCminus07mm

Wild

-type

ApoE

4

tractOptic

02

04

06

08

Frac

tiona

l ani

sotro

py (F

A)

(c)





8 Neural Plasticity

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

Fortasyn

Control

Fortasyn

ApoE

4

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Motor cortex

SS cortex

Visual cortex

Dorsal Hipp

Ventral Hipp

Auditory cortex

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp

12-month-oldControl

Wild

-type


Ventral Hipp

Auditory cortex

Motor cortex

Dorsal Hipp

SS cortex

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

Wild

-type

ApoE

4

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

Fortasyn

Control

Fortasyn

18-month-oldControl

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Motor cortex

SS cortex

Visual cortex

Dorsal Hipp

Ventral Hipp

Auditory cortex

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


SS cortex

Visual cortex

Ventral Hipp

Auditory cortex

Motor cortex

Dorsal Hipp

ApoE

4W

ild-ty

peW

ild-ty

peAp

oE4






Neural Plasticity 9

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

12-month-old

Control

Fortasyn

Control

Fortasyn

ApoE

4W

ild-ty

pe

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Dorsal Hipp

Motor cortex

SS cortex

Visual cortex

Ventral Hipp

Auditory cortex


Ventral Hipp

Auditory cortex

Motor cortex

Visual cortex

Dorsal Hipp

SS cortex

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

Wild

-type

ApoE

4

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

18-month-old

Control

Fortasyn

Control

Fortasyn

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Dorsal Hipp

Motor cortex

SS cortex

Visual cortex

Ventral Hipp

Auditory cortex


Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex

Dorsal Hipp

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp

ApoE

4W

ild-ty

peW

ild-ty

peAp

oE4



12-month-old

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

Visualcortex

Sensorycortex

IML(DG)(DG)


CA3

0

50000

100000

150000

PSD

-95

stai

ned

area

ControlFortasyn

(a)

lowast lowastlowast

lowastlowast

lowast

lowast

18-month-old

Visualcortex

Sensorycortex

IML(DG)(DG)


CA3

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

0

50000

100000

150000

PSD

-95

stain

ed ar

ea

ControlFortasyn

(b)






4 Discussion















5 Conclusions




Abbreviations





Competing Interests




Acknowledgments


References






































































































Neural Plasticity










Psychiatry Journal









Journal of


2 Neural Plasticity










Neural Plasticity 3



Control[g100 g]

Fortasyn[g100 g]


2


BirthStart diets

0 2 4 10 12 14 16 18


Months






2








4 Neural Plasticity








3 Results


31 Body Weight




Neural Plasticity 5

ControlFortasyn

lowast

lowastlowast


lowastlowastlowast



0

10

20

30

40

50

60

70Bo

dy w

eigh

t (g)

(a)

ControlFortasyn

lowast

lowastlowast

lowast

lowastlowast



0

10

20

30

40

50

60

70

Body

wei

ght (

g)

(b)












323 rsfMRI




6 Neural Plasticity

12-month-oldlowast

0

100

200

300



Wild-type

Wild-type

CBF

(mL100

gm

in)

ControlFortasyn

(a)

0

100

200

300


18-month-old

lowast lowast

lowast


Wild-type

Wild-type

CBF

(mL100

gm

in)

ControlFortasyn

lowast

(b)












MD


12-month-old

ApoE

4W

ild-ty

pe

tractOptic

Wild

-type

ApoE

4

CCminus07mm

Wild

-type

ApoE

4

CCminus17mm

ApoE

4W

ild-ty

pe

CCminus22mm

ApoE

4W

ild-ty

pe

Fornix

02

04

06

08

Frac

tiona

l ani

sotro

py (F

A)

lowastlowast

ControlFortasyn

lowast

lowastlowast

00006

00007

00008

00009

00010

Mea

n di

ffusiv

ity (M

D) (

mm

2s

)

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

Motorcortex

Auditorycortex

Sensorycortex

Visualcortex

ApoE

4W

ild-ty

pe

Hippocampus

(b)

ControlFortasyn

lowastlowast

00006

00007

00008

00009

00010

Mea

n di

ffusiv

ity (M

D) (

mm

2s

)

Wild

-type

ApoE

4

Motorcortex

Wild

-type

ApoE

4

Auditorycortex

Wild

-type

ApoE

4

Sensorycortex

Wild

-type

ApoE

4Visualcortex

Wild

-type

ApoE

4

Hippocampus

18-month-old

lowastlowast

ApoE

4W

ild-ty

pe

Fornix

Wild

-type

ApoE

4

CCminus22mm

Wild

-type

ApoE

4

CCminus17mm

Wild

-type

ApoE

4

CCminus07mm

Wild

-type

ApoE

4

tractOptic

02

04

06

08

Frac

tiona

l ani

sotro

py (F

A)

(c)





8 Neural Plasticity

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

Fortasyn

Control

Fortasyn

ApoE

4

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Motor cortex

SS cortex

Visual cortex

Dorsal Hipp

Ventral Hipp

Auditory cortex

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp

12-month-oldControl

Wild

-type


Ventral Hipp

Auditory cortex

Motor cortex

Dorsal Hipp

SS cortex

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

Wild

-type

ApoE

4

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

Fortasyn

Control

Fortasyn

18-month-oldControl

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Motor cortex

SS cortex

Visual cortex

Dorsal Hipp

Ventral Hipp

Auditory cortex

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


SS cortex

Visual cortex

Ventral Hipp

Auditory cortex

Motor cortex

Dorsal Hipp

ApoE

4W

ild-ty

peW

ild-ty

peAp

oE4






Neural Plasticity 9

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

12-month-old

Control

Fortasyn

Control

Fortasyn

ApoE

4W

ild-ty

pe

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Dorsal Hipp

Motor cortex

SS cortex

Visual cortex

Ventral Hipp

Auditory cortex


Ventral Hipp

Auditory cortex

Motor cortex

Visual cortex

Dorsal Hipp

SS cortex

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

Wild

-type

ApoE

4

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

18-month-old

Control

Fortasyn

Control

Fortasyn

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Dorsal Hipp

Motor cortex

SS cortex

Visual cortex

Ventral Hipp

Auditory cortex


Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex

Dorsal Hipp

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp

ApoE

4W

ild-ty

peW

ild-ty

peAp

oE4



12-month-old

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

Visualcortex

Sensorycortex

IML(DG)(DG)


CA3

0

50000

100000

150000

PSD

-95

stai

ned

area

ControlFortasyn

(a)

lowast lowastlowast

lowastlowast

lowast

lowast

18-month-old

Visualcortex

Sensorycortex

IML(DG)(DG)


CA3

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

0

50000

100000

150000

PSD

-95

stain

ed ar

ea

ControlFortasyn

(b)






4 Discussion















5 Conclusions




Abbreviations





Competing Interests




Acknowledgments


References






































































































Neural Plasticity










Psychiatry Journal









Journal of


Neural Plasticity 3



Control[g100 g]

Fortasyn[g100 g]


2


BirthStart diets

0 2 4 10 12 14 16 18


Months






2








4 Neural Plasticity








3 Results


31 Body Weight




Neural Plasticity 5

ControlFortasyn

lowast

lowastlowast


lowastlowastlowast



0

10

20

30

40

50

60

70Bo

dy w

eigh

t (g)

(a)

ControlFortasyn

lowast

lowastlowast

lowast

lowastlowast



0

10

20

30

40

50

60

70

Body

wei

ght (

g)

(b)












323 rsfMRI




6 Neural Plasticity

12-month-oldlowast

0

100

200

300



Wild-type

Wild-type

CBF

(mL100

gm

in)

ControlFortasyn

(a)

0

100

200

300


18-month-old

lowast lowast

lowast


Wild-type

Wild-type

CBF

(mL100

gm

in)

ControlFortasyn

lowast

(b)












MD


12-month-old

ApoE

4W

ild-ty

pe

tractOptic

Wild

-type

ApoE

4

CCminus07mm

Wild

-type

ApoE

4

CCminus17mm

ApoE

4W

ild-ty

pe

CCminus22mm

ApoE

4W

ild-ty

pe

Fornix

02

04

06

08

Frac

tiona

l ani

sotro

py (F

A)

lowastlowast

ControlFortasyn

lowast

lowastlowast

00006

00007

00008

00009

00010

Mea

n di

ffusiv

ity (M

D) (

mm

2s

)

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

Motorcortex

Auditorycortex

Sensorycortex

Visualcortex

ApoE

4W

ild-ty

pe

Hippocampus

(b)

ControlFortasyn

lowastlowast

00006

00007

00008

00009

00010

Mea

n di

ffusiv

ity (M

D) (

mm

2s

)

Wild

-type

ApoE

4

Motorcortex

Wild

-type

ApoE

4

Auditorycortex

Wild

-type

ApoE

4

Sensorycortex

Wild

-type

ApoE

4Visualcortex

Wild

-type

ApoE

4

Hippocampus

18-month-old

lowastlowast

ApoE

4W

ild-ty

pe

Fornix

Wild

-type

ApoE

4

CCminus22mm

Wild

-type

ApoE

4

CCminus17mm

Wild

-type

ApoE

4

CCminus07mm

Wild

-type

ApoE

4

tractOptic

02

04

06

08

Frac

tiona

l ani

sotro

py (F

A)

(c)





8 Neural Plasticity

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

Fortasyn

Control

Fortasyn

ApoE

4

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Motor cortex

SS cortex

Visual cortex

Dorsal Hipp

Ventral Hipp

Auditory cortex

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp

12-month-oldControl

Wild

-type


Ventral Hipp

Auditory cortex

Motor cortex

Dorsal Hipp

SS cortex

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

Wild

-type

ApoE

4

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

Fortasyn

Control

Fortasyn

18-month-oldControl

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Motor cortex

SS cortex

Visual cortex

Dorsal Hipp

Ventral Hipp

Auditory cortex

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


SS cortex

Visual cortex

Ventral Hipp

Auditory cortex

Motor cortex

Dorsal Hipp

ApoE

4W

ild-ty

peW

ild-ty

peAp

oE4






Neural Plasticity 9

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

12-month-old

Control

Fortasyn

Control

Fortasyn

ApoE

4W

ild-ty

pe

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Dorsal Hipp

Motor cortex

SS cortex

Visual cortex

Ventral Hipp

Auditory cortex


Ventral Hipp

Auditory cortex

Motor cortex

Visual cortex

Dorsal Hipp

SS cortex

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

Wild

-type

ApoE

4

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

18-month-old

Control

Fortasyn

Control

Fortasyn

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Dorsal Hipp

Motor cortex

SS cortex

Visual cortex

Ventral Hipp

Auditory cortex


Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex

Dorsal Hipp

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp

ApoE

4W

ild-ty

peW

ild-ty

peAp

oE4



12-month-old

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

Visualcortex

Sensorycortex

IML(DG)(DG)


CA3

0

50000

100000

150000

PSD

-95

stai

ned

area

ControlFortasyn

(a)

lowast lowastlowast

lowastlowast

lowast

lowast

18-month-old

Visualcortex

Sensorycortex

IML(DG)(DG)


CA3

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

0

50000

100000

150000

PSD

-95

stain

ed ar

ea

ControlFortasyn

(b)






4 Discussion















5 Conclusions




Abbreviations





Competing Interests




Acknowledgments


References






































































































Neural Plasticity










Psychiatry Journal









Journal of


4 Neural Plasticity








3 Results


31 Body Weight




Neural Plasticity 5

ControlFortasyn

lowast

lowastlowast


lowastlowastlowast



0

10

20

30

40

50

60

70Bo

dy w

eigh

t (g)

(a)

ControlFortasyn

lowast

lowastlowast

lowast

lowastlowast



0

10

20

30

40

50

60

70

Body

wei

ght (

g)

(b)












323 rsfMRI




6 Neural Plasticity

12-month-oldlowast

0

100

200

300



Wild-type

Wild-type

CBF

(mL100

gm

in)

ControlFortasyn

(a)

0

100

200

300


18-month-old

lowast lowast

lowast


Wild-type

Wild-type

CBF

(mL100

gm

in)

ControlFortasyn

lowast

(b)












MD


12-month-old

ApoE

4W

ild-ty

pe

tractOptic

Wild

-type

ApoE

4

CCminus07mm

Wild

-type

ApoE

4

CCminus17mm

ApoE

4W

ild-ty

pe

CCminus22mm

ApoE

4W

ild-ty

pe

Fornix

02

04

06

08

Frac

tiona

l ani

sotro

py (F

A)

lowastlowast

ControlFortasyn

lowast

lowastlowast

00006

00007

00008

00009

00010

Mea

n di

ffusiv

ity (M

D) (

mm

2s

)

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

Motorcortex

Auditorycortex

Sensorycortex

Visualcortex

ApoE

4W

ild-ty

pe

Hippocampus

(b)

ControlFortasyn

lowastlowast

00006

00007

00008

00009

00010

Mea

n di

ffusiv

ity (M

D) (

mm

2s

)

Wild

-type

ApoE

4

Motorcortex

Wild

-type

ApoE

4

Auditorycortex

Wild

-type

ApoE

4

Sensorycortex

Wild

-type

ApoE

4Visualcortex

Wild

-type

ApoE

4

Hippocampus

18-month-old

lowastlowast

ApoE

4W

ild-ty

pe

Fornix

Wild

-type

ApoE

4

CCminus22mm

Wild

-type

ApoE

4

CCminus17mm

Wild

-type

ApoE

4

CCminus07mm

Wild

-type

ApoE

4

tractOptic

02

04

06

08

Frac

tiona

l ani

sotro

py (F

A)

(c)





8 Neural Plasticity

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

Fortasyn

Control

Fortasyn

ApoE

4

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Motor cortex

SS cortex

Visual cortex

Dorsal Hipp

Ventral Hipp

Auditory cortex

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp

12-month-oldControl

Wild

-type


Ventral Hipp

Auditory cortex

Motor cortex

Dorsal Hipp

SS cortex

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

Wild

-type

ApoE

4

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

Fortasyn

Control

Fortasyn

18-month-oldControl

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Motor cortex

SS cortex

Visual cortex

Dorsal Hipp

Ventral Hipp

Auditory cortex

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


SS cortex

Visual cortex

Ventral Hipp

Auditory cortex

Motor cortex

Dorsal Hipp

ApoE

4W

ild-ty

peW

ild-ty

peAp

oE4






Neural Plasticity 9

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

12-month-old

Control

Fortasyn

Control

Fortasyn

ApoE

4W

ild-ty

pe

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Dorsal Hipp

Motor cortex

SS cortex

Visual cortex

Ventral Hipp

Auditory cortex


Ventral Hipp

Auditory cortex

Motor cortex

Visual cortex

Dorsal Hipp

SS cortex

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

Wild

-type

ApoE

4

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

18-month-old

Control

Fortasyn

Control

Fortasyn

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Dorsal Hipp

Motor cortex

SS cortex

Visual cortex

Ventral Hipp

Auditory cortex


Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex

Dorsal Hipp

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp

ApoE

4W

ild-ty

peW

ild-ty

peAp

oE4



12-month-old

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

Visualcortex

Sensorycortex

IML(DG)(DG)


CA3

0

50000

100000

150000

PSD

-95

stai

ned

area

ControlFortasyn

(a)

lowast lowastlowast

lowastlowast

lowast

lowast

18-month-old

Visualcortex

Sensorycortex

IML(DG)(DG)


CA3

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

0

50000

100000

150000

PSD

-95

stain

ed ar

ea

ControlFortasyn

(b)






4 Discussion















5 Conclusions




Abbreviations





Competing Interests




Acknowledgments


References






































































































Neural Plasticity










Psychiatry Journal









Journal of


Neural Plasticity 5

ControlFortasyn

lowast

lowastlowast


lowastlowastlowast



0

10

20

30

40

50

60

70Bo

dy w

eigh

t (g)

(a)

ControlFortasyn

lowast

lowastlowast

lowast

lowastlowast



0

10

20

30

40

50

60

70

Body

wei

ght (

g)

(b)












323 rsfMRI




6 Neural Plasticity

12-month-oldlowast

0

100

200

300



Wild-type

Wild-type

CBF

(mL100

gm

in)

ControlFortasyn

(a)

0

100

200

300


18-month-old

lowast lowast

lowast


Wild-type

Wild-type

CBF

(mL100

gm

in)

ControlFortasyn

lowast

(b)












MD


12-month-old

ApoE

4W

ild-ty

pe

tractOptic

Wild

-type

ApoE

4

CCminus07mm

Wild

-type

ApoE

4

CCminus17mm

ApoE

4W

ild-ty

pe

CCminus22mm

ApoE

4W

ild-ty

pe

Fornix

02

04

06

08

Frac

tiona

l ani

sotro

py (F

A)

lowastlowast

ControlFortasyn

lowast

lowastlowast

00006

00007

00008

00009

00010

Mea

n di

ffusiv

ity (M

D) (

mm

2s

)

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

Motorcortex

Auditorycortex

Sensorycortex

Visualcortex

ApoE

4W

ild-ty

pe

Hippocampus

(b)

ControlFortasyn

lowastlowast

00006

00007

00008

00009

00010

Mea

n di

ffusiv

ity (M

D) (

mm

2s

)

Wild

-type

ApoE

4

Motorcortex

Wild

-type

ApoE

4

Auditorycortex

Wild

-type

ApoE

4

Sensorycortex

Wild

-type

ApoE

4Visualcortex

Wild

-type

ApoE

4

Hippocampus

18-month-old

lowastlowast

ApoE

4W

ild-ty

pe

Fornix

Wild

-type

ApoE

4

CCminus22mm

Wild

-type

ApoE

4

CCminus17mm

Wild

-type

ApoE

4

CCminus07mm

Wild

-type

ApoE

4

tractOptic

02

04

06

08

Frac

tiona

l ani

sotro

py (F

A)

(c)





8 Neural Plasticity

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

Fortasyn

Control

Fortasyn

ApoE

4

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Motor cortex

SS cortex

Visual cortex

Dorsal Hipp

Ventral Hipp

Auditory cortex

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp

12-month-oldControl

Wild

-type


Ventral Hipp

Auditory cortex

Motor cortex

Dorsal Hipp

SS cortex

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

Wild

-type

ApoE

4

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

Fortasyn

Control

Fortasyn

18-month-oldControl

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Motor cortex

SS cortex

Visual cortex

Dorsal Hipp

Ventral Hipp

Auditory cortex

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


SS cortex

Visual cortex

Ventral Hipp

Auditory cortex

Motor cortex

Dorsal Hipp

ApoE

4W

ild-ty

peW

ild-ty

peAp

oE4






Neural Plasticity 9

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

12-month-old

Control

Fortasyn

Control

Fortasyn

ApoE

4W

ild-ty

pe

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Dorsal Hipp

Motor cortex

SS cortex

Visual cortex

Ventral Hipp

Auditory cortex


Ventral Hipp

Auditory cortex

Motor cortex

Visual cortex

Dorsal Hipp

SS cortex

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

Wild

-type

ApoE

4

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

18-month-old

Control

Fortasyn

Control

Fortasyn

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Dorsal Hipp

Motor cortex

SS cortex

Visual cortex

Ventral Hipp

Auditory cortex


Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex

Dorsal Hipp

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp

ApoE

4W

ild-ty

peW

ild-ty

peAp

oE4



12-month-old

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

Visualcortex

Sensorycortex

IML(DG)(DG)


CA3

0

50000

100000

150000

PSD

-95

stai

ned

area

ControlFortasyn

(a)

lowast lowastlowast

lowastlowast

lowast

lowast

18-month-old

Visualcortex

Sensorycortex

IML(DG)(DG)


CA3

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

0

50000

100000

150000

PSD

-95

stain

ed ar

ea

ControlFortasyn

(b)






4 Discussion















5 Conclusions




Abbreviations





Competing Interests




Acknowledgments


References






































































































Neural Plasticity










Psychiatry Journal









Journal of


6 Neural Plasticity

12-month-oldlowast

0

100

200

300



Wild-type

Wild-type

CBF

(mL100

gm

in)

ControlFortasyn

(a)

0

100

200

300


18-month-old

lowast lowast

lowast


Wild-type

Wild-type

CBF

(mL100

gm

in)

ControlFortasyn

lowast

(b)












MD


12-month-old

ApoE

4W

ild-ty

pe

tractOptic

Wild

-type

ApoE

4

CCminus07mm

Wild

-type

ApoE

4

CCminus17mm

ApoE

4W

ild-ty

pe

CCminus22mm

ApoE

4W

ild-ty

pe

Fornix

02

04

06

08

Frac

tiona

l ani

sotro

py (F

A)

lowastlowast

ControlFortasyn

lowast

lowastlowast

00006

00007

00008

00009

00010

Mea

n di

ffusiv

ity (M

D) (

mm

2s

)

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

Motorcortex

Auditorycortex

Sensorycortex

Visualcortex

ApoE

4W

ild-ty

pe

Hippocampus

(b)

ControlFortasyn

lowastlowast

00006

00007

00008

00009

00010

Mea

n di

ffusiv

ity (M

D) (

mm

2s

)

Wild

-type

ApoE

4

Motorcortex

Wild

-type

ApoE

4

Auditorycortex

Wild

-type

ApoE

4

Sensorycortex

Wild

-type

ApoE

4Visualcortex

Wild

-type

ApoE

4

Hippocampus

18-month-old

lowastlowast

ApoE

4W

ild-ty

pe

Fornix

Wild

-type

ApoE

4

CCminus22mm

Wild

-type

ApoE

4

CCminus17mm

Wild

-type

ApoE

4

CCminus07mm

Wild

-type

ApoE

4

tractOptic

02

04

06

08

Frac

tiona

l ani

sotro

py (F

A)

(c)





8 Neural Plasticity

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

Fortasyn

Control

Fortasyn

ApoE

4

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Motor cortex

SS cortex

Visual cortex

Dorsal Hipp

Ventral Hipp

Auditory cortex

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp

12-month-oldControl

Wild

-type


Ventral Hipp

Auditory cortex

Motor cortex

Dorsal Hipp

SS cortex

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

Wild

-type

ApoE

4

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

Fortasyn

Control

Fortasyn

18-month-oldControl

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Motor cortex

SS cortex

Visual cortex

Dorsal Hipp

Ventral Hipp

Auditory cortex

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


SS cortex

Visual cortex

Ventral Hipp

Auditory cortex

Motor cortex

Dorsal Hipp

ApoE

4W

ild-ty

peW

ild-ty

peAp

oE4






Neural Plasticity 9

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

12-month-old

Control

Fortasyn

Control

Fortasyn

ApoE

4W

ild-ty

pe

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Dorsal Hipp

Motor cortex

SS cortex

Visual cortex

Ventral Hipp

Auditory cortex


Ventral Hipp

Auditory cortex

Motor cortex

Visual cortex

Dorsal Hipp

SS cortex

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

Wild

-type

ApoE

4

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

18-month-old

Control

Fortasyn

Control

Fortasyn

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Dorsal Hipp

Motor cortex

SS cortex

Visual cortex

Ventral Hipp

Auditory cortex


Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex

Dorsal Hipp

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp

ApoE

4W

ild-ty

peW

ild-ty

peAp

oE4



12-month-old

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

Visualcortex

Sensorycortex

IML(DG)(DG)


CA3

0

50000

100000

150000

PSD

-95

stai

ned

area

ControlFortasyn

(a)

lowast lowastlowast

lowastlowast

lowast

lowast

18-month-old

Visualcortex

Sensorycortex

IML(DG)(DG)


CA3

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

0

50000

100000

150000

PSD

-95

stain

ed ar

ea

ControlFortasyn

(b)






4 Discussion















5 Conclusions




Abbreviations





Competing Interests




Acknowledgments


References






































































































Neural Plasticity










Psychiatry Journal









Journal of



MD


12-month-old

ApoE

4W

ild-ty

pe

tractOptic

Wild

-type

ApoE

4

CCminus07mm

Wild

-type

ApoE

4

CCminus17mm

ApoE

4W

ild-ty

pe

CCminus22mm

ApoE

4W

ild-ty

pe

Fornix

02

04

06

08

Frac

tiona

l ani

sotro

py (F

A)

lowastlowast

ControlFortasyn

lowast

lowastlowast

00006

00007

00008

00009

00010

Mea

n di

ffusiv

ity (M

D) (

mm

2s

)

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

Motorcortex

Auditorycortex

Sensorycortex

Visualcortex

ApoE

4W

ild-ty

pe

Hippocampus

(b)

ControlFortasyn

lowastlowast

00006

00007

00008

00009

00010

Mea

n di

ffusiv

ity (M

D) (

mm

2s

)

Wild

-type

ApoE

4

Motorcortex

Wild

-type

ApoE

4

Auditorycortex

Wild

-type

ApoE

4

Sensorycortex

Wild

-type

ApoE

4Visualcortex

Wild

-type

ApoE

4

Hippocampus

18-month-old

lowastlowast

ApoE

4W

ild-ty

pe

Fornix

Wild

-type

ApoE

4

CCminus22mm

Wild

-type

ApoE

4

CCminus17mm

Wild

-type

ApoE

4

CCminus07mm

Wild

-type

ApoE

4

tractOptic

02

04

06

08

Frac

tiona

l ani

sotro

py (F

A)

(c)





8 Neural Plasticity

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

Fortasyn

Control

Fortasyn

ApoE

4

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Motor cortex

SS cortex

Visual cortex

Dorsal Hipp

Ventral Hipp

Auditory cortex

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp

12-month-oldControl

Wild

-type


Ventral Hipp

Auditory cortex

Motor cortex

Dorsal Hipp

SS cortex

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

Wild

-type

ApoE

4

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

Fortasyn

Control

Fortasyn

18-month-oldControl

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Motor cortex

SS cortex

Visual cortex

Dorsal Hipp

Ventral Hipp

Auditory cortex

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


SS cortex

Visual cortex

Ventral Hipp

Auditory cortex

Motor cortex

Dorsal Hipp

ApoE

4W

ild-ty

peW

ild-ty

peAp

oE4






Neural Plasticity 9

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

12-month-old

Control

Fortasyn

Control

Fortasyn

ApoE

4W

ild-ty

pe

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Dorsal Hipp

Motor cortex

SS cortex

Visual cortex

Ventral Hipp

Auditory cortex


Ventral Hipp

Auditory cortex

Motor cortex

Visual cortex

Dorsal Hipp

SS cortex

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

Wild

-type

ApoE

4

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

18-month-old

Control

Fortasyn

Control

Fortasyn

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Dorsal Hipp

Motor cortex

SS cortex

Visual cortex

Ventral Hipp

Auditory cortex


Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex

Dorsal Hipp

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp

ApoE

4W

ild-ty

peW

ild-ty

peAp

oE4



12-month-old

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

Visualcortex

Sensorycortex

IML(DG)(DG)


CA3

0

50000

100000

150000

PSD

-95

stai

ned

area

ControlFortasyn

(a)

lowast lowastlowast

lowastlowast

lowast

lowast

18-month-old

Visualcortex

Sensorycortex

IML(DG)(DG)


CA3

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

0

50000

100000

150000

PSD

-95

stain

ed ar

ea

ControlFortasyn

(b)






4 Discussion















5 Conclusions




Abbreviations





Competing Interests




Acknowledgments


References






































































































Neural Plasticity










Psychiatry Journal









Journal of


8 Neural Plasticity

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

Fortasyn

Control

Fortasyn

ApoE

4

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Motor cortex

SS cortex

Visual cortex

Dorsal Hipp

Ventral Hipp

Auditory cortex

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp

12-month-oldControl

Wild

-type


Ventral Hipp

Auditory cortex

Motor cortex

Dorsal Hipp

SS cortex

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

Wild

-type

ApoE

4

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

Fortasyn

Control

Fortasyn

18-month-oldControl

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

z-s

core

30

40

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Motor cortex

SS cortex

Visual cortex

Dorsal Hipp

Ventral Hipp

Auditory cortex

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


SS cortex

Visual cortex

Ventral Hipp

Auditory cortex

Motor cortex

Dorsal Hipp

ApoE

4W

ild-ty

peW

ild-ty

peAp

oE4






Neural Plasticity 9

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

12-month-old

Control

Fortasyn

Control

Fortasyn

ApoE

4W

ild-ty

pe

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Dorsal Hipp

Motor cortex

SS cortex

Visual cortex

Ventral Hipp

Auditory cortex


Ventral Hipp

Auditory cortex

Motor cortex

Visual cortex

Dorsal Hipp

SS cortex

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

Wild

-type

ApoE

4

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

18-month-old

Control

Fortasyn

Control

Fortasyn

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Dorsal Hipp

Motor cortex

SS cortex

Visual cortex

Ventral Hipp

Auditory cortex


Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex

Dorsal Hipp

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp

ApoE

4W

ild-ty

peW

ild-ty

peAp

oE4



12-month-old

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

Visualcortex

Sensorycortex

IML(DG)(DG)


CA3

0

50000

100000

150000

PSD

-95

stai

ned

area

ControlFortasyn

(a)

lowast lowastlowast

lowastlowast

lowast

lowast

18-month-old

Visualcortex

Sensorycortex

IML(DG)(DG)


CA3

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

0

50000

100000

150000

PSD

-95

stain

ed ar

ea

ControlFortasyn

(b)






4 Discussion















5 Conclusions




Abbreviations





Competing Interests




Acknowledgments


References






































































































Neural Plasticity










Psychiatry Journal









Journal of


Neural Plasticity 9

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

12-month-old

Control

Fortasyn

Control

Fortasyn

ApoE

4W

ild-ty

pe

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Dorsal Hipp

Motor cortex

SS cortex

Visual cortex

Ventral Hipp

Auditory cortex


Ventral Hipp

Auditory cortex

Motor cortex

Visual cortex

Dorsal Hipp

SS cortex

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

Wild

-type

ApoE

4

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

LRLRLRLRLRLR

18-month-old

Control

Fortasyn

Control

Fortasyn

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

196

z-s

core

10

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp


Dorsal Hipp

Motor cortex

SS cortex

Visual cortex

Ventral Hipp

Auditory cortex


Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex

Dorsal Hipp

Ventral Hipp

Auditory cortex

Motor cortex

SS cortex

Visual cortex


Dorsal Hipp

ApoE

4W

ild-ty

peW

ild-ty

peAp

oE4



12-month-old

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

Visualcortex

Sensorycortex

IML(DG)(DG)


CA3

0

50000

100000

150000

PSD

-95

stai

ned

area

ControlFortasyn

(a)

lowast lowastlowast

lowastlowast

lowast

lowast

18-month-old

Visualcortex

Sensorycortex

IML(DG)(DG)


CA3

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

0

50000

100000

150000

PSD

-95

stain

ed ar

ea

ControlFortasyn

(b)






4 Discussion















5 Conclusions




Abbreviations





Competing Interests




Acknowledgments


References






































































































Neural Plasticity










Psychiatry Journal









Journal of



12-month-old

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

Visualcortex

Sensorycortex

IML(DG)(DG)


CA3

0

50000

100000

150000

PSD

-95

stai

ned

area

ControlFortasyn

(a)

lowast lowastlowast

lowastlowast

lowast

lowast

18-month-old

Visualcortex

Sensorycortex

IML(DG)(DG)


CA3

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

ApoE

4W

ild-ty

pe

0

50000

100000

150000

PSD

-95

stain

ed ar

ea

ControlFortasyn

(b)






4 Discussion















5 Conclusions




Abbreviations





Competing Interests




Acknowledgments


References






































































































Neural Plasticity










Psychiatry Journal









Journal of










5 Conclusions




Abbreviations





Competing Interests




Acknowledgments


References






































































































Neural Plasticity










Psychiatry Journal









Journal of




Abbreviations





Competing Interests




Acknowledgments


References






































































































Neural Plasticity










Psychiatry Journal









Journal of































































































Neural Plasticity










Psychiatry Journal









Journal of































































Neural Plasticity










Psychiatry Journal









Journal of




























Neural Plasticity










Psychiatry Journal









Journal of














Neural Plasticity










Psychiatry Journal









Journal of


Documents

PDF hosted at the Radboud Repository of the Radboud University … · 2017-12-06 · Research Article A Dietary Treatment Improves Cerebral Blood Flow and Brain Connectivity in Aging