DAG IP3

Молекулярные и клеточные механизмы обоняния

Olfaction — a mode of communication between animals

Recognition

stress signals (predator, aggression, fear)food identification territorial boundaries

Reproductive behavior

mate selectioncourtshipsexual behavior, maternal behaviorkin recognition

Brain

MOEVNO

Septal

organ

Grueneberg

ganglion

MOB

AOB

Complexity of the mammalian olfactory system

Septal organ

Glueneberg ganglion

Vomeronasal organ

Main

olfactory

epithelium

Main

olfactory

bulb

Accessory

olfactory

bulb

BV

basal

apical

ethyl octanoate4-heptanolide

Minty

Beta-cyclocitral

p-anisaldegyde 2,6-dimethylpyrazine

Fruity Nutty

Odorant space

General odorants are small hydrophobic molecules

volatile at ambient temperature

Tree types of cone photoreceptors

D-камфора

Борнеол

Цинеол

Амилмеркаптан

Пентаметилендиамин

camphor odor putrefactive odor

bitter almond odor

ferrocene nickelocene

Fe Ni

spicy odor oily odor

benzaldehyde hydrogene cyanide

CH3-CH2-CH2-CH2-CH2-CH2-C-OCH3||

O

CH3-CH2-CH2-CH2-CH2-CH2-C-H||

O

CH3-CH2-CH2-CH2-CH2-CH2-C-CH3||

O

CH3-CH2-CH2-CH2-CH2-CH2-C-OH||

O

1-heptanol - citrus, sweet

1-heptanal - oily, unplesunt

Methyl heptanoate – pineapple, sweet

2-octanone – sweet, non-citrus

1-heptanoic acid – paint, chemical

Functional group theory

CH3-CH2-CH2-CH2-CH2-CH2-C-OH|

|

H

H

(4S,7R)-(-)-galaxolide

powerful musk odor(4R,7S)-(+)-galaxolide

almost odorless

(3R)-(+)-Fixolide

light, sweet, aromatic

(3S)-(-)-Fixolide

strong musk

1-heptanol

Laing et al., Chemical Senses 2003

John Earnest Amoore

(1930-1998)

Isovaleric acid

The stereochemical theory for olfaction

The Nobel Prize in Physiology and Medicine 2004 for the discovery of

odorant receptors

Linda BuckRichard Axel

Species Number of OR genes

Man 396

Chimpanzee 399

Macaque 326

Mouse 1130

Rat 1207

Dog 811

Caw 1152

Opossum 1198

Chicken 300

Xenopus 1024

Zebra fish 155

Fugu 86

Drosophila 62

C.elegans 1278

Receptors operative in MOE

Receptors Ligands Origin Proposed functions

odorantGPCRs

N = 350-1300

trace amineGPCRN = 3-20

GC-D

N = 1

general odors

MHC class I

peptides

volatile amines

CO2

uroguanyline

guanylin

food

environment

urine

body secretions

urine

atmosphere

urine

odor recognition, discrimination,

attraction, repulsion

social recognition of

other strains

stress response

gender recognition

avoidance behavior

Detection of cues related to

hunger, satiety, or thirst

Olfactory transduction

Vt

Vt = SaiNiYi

Distinct cell subpopulations contribute additively to electroolfactogram

SC

OSN

Responses of the main olfactory epithelium to different odorants

Human olfactory sensory neurons

Images by Constanze R.M. and Morrison E.E., Virginia University

Boekhoff, Breer, PNAS 1992

cAMP generation in olfactory cilia

(A) Olfactory sensory neuron

(B) Supporting cell

Curr

ent (p

A)

Time (s) Lilial (mM)

Pe

ak c

urr

en

t (p

A)

lilial

Electrical excitation of olfactory neurons by odorants

Patch clamp method for recording channel activity

Amplifier

Recordingpipette

Assayed cell

50 ms

50

5

2

0.5

0.1

0

cA

MP

-gate

d c

urr

ent

(pA

)

cAMP,m

-6

-5

-6

-4

-2

-6

-4

-2

-4

-2

0

-1

0

-1

0

AC transduction cascade in olfactory cilia

PDE1

ACIII

Odorant

Ca2+

Cl-

3Na+

1Ca2+

abg

Receptor

CaM

cAMP-gated channel

Arrestin

RKGolf

cAMP

CNGA2

PDE4

1Ca2+1Na+/ 2Cl-/1K+1K+

4Na+

Cl-in ~ 40-100 mM

Cl-out ~ 55-93 mM

Adaptation

stimulus

binding transduction response

Adaptation machinery

Control response

Adapted response

Definition of cellular adaptation

Cineol, 300 mM

50 pA

1 sec

Cineol, 300 mM

3 mM Ca2+

3 mM Mg2+

, no Ca2+

A

B

Kurahashi & Shibuya, Brain Res., 1990

Adaptation of olfactory sensory neurons is mediated by Ca2+

Ca2+-dependent elements of the olfactory transduction cascade

PDE1

ACIII

Odorant

Ca2+

Cl-

3Na+

1Ca2+

abg

Receptor

CaM

cAMP-gated channel

Arrestin

RKGolf

cAMP

CNGA2

1Ca2+1Na+/ 2Cl-/1K+1K+

4Na+

CNGA2

CNGA4

CNGB1b

CaM siteCNBD

Olfactory CNG channel

After Trudeau & Zagotta, J.Biol.Chem. 2003

CNGA2CNGA2

CNGB1b CNGA4

CaM site

0.01 0.1 1 10 100 1000 10000

0.0

0.5

1.0

cAMP concentration, mM

Norm

aliz

ed o

pe

n p

rob

ab

ility

K1/2 = 1.4 mM K1/2 = 29 mM

Control Calmodulin

Odorant

receptors

Trace amine

receptors

Golf

ACIII

CNG2A

PDE4

Ciliary OMP-positive

Microvillar

OMP-negative

Receptor-GC

CNGA3

PDE2

?

TRPM5 gustducin

PLCb2

TRPM5

?

Population of chemosensory cells in MOE

GC-cGMP transduction cascade in olfactory cilia

PDE2

Guanylyl

cyclase-D

cGMP-gated channel

cGMP

CNGA3

DF

/F=

10%

50 s

0.4%

Hu et al., Science 2007

OSN

Leinders-Zufall et al., PNAS 2007

uroguanylin guanylin

Species Functionality of GC-D gene

Mouse +

Rat +

Treeshrew +

Lemur +

bushbaby +

Man -

Chimpanzee -

Macaque -

Orangutan -

Gorilla -

Baboon -

Encoding

OSN responses to different odorants

Furudono et al. Chem Senses, 2009.

Agonist types

0.1 1 10 100 1000 10000

0

1

Ligand concentration

Norm

aliz

ed

resp

onse

Full agonist

Partial agonist

Inverse agonist

Antagonist

Pro

babili

ty

Conformation

basal full agonist

partial agonist

inverse agonist

acetyl isoeugenol (1)

eugenol (EG) methyl isoeugenol (2)

allyl benzene (3)

isosafrole (4)

Odorant antagonism

receptors

odorant 2odorant 1 mixture

ORs

code

ORs

Odorant coding

Vassalli et al., Cell, 2002

Axons of M71-espressing OSN converge into two glomeruli

glomeruli

axons

Olfactory sensory

neurons

M71-LacZ transgenic mouse

The discrete nature of the matter underlies the Schottky noise

A ~ N D – (A-A)2~ N = D

A = /A ~ 1/N

-50 mV

10 mV

20 ms

1 pA N = 280

Vomeronasal organ

Septal organ

Glueneberg ganglion

Vomeronasal organ

Main

olfactory

epithelium

Main

olfactory

bulb

Accessory

olfactory

bulb

BV

basal

apical

Compound Species Effect Olfactory subsystem

3-Amino-s-triazole Mouse Attracts females ?

2,5-Dimethylpyrazine Mouse Delays puberty VNO

in females

2-Heptanone Mouse Extends estrus VNO

4-Ethylphenol Mouse Attracts females and VNO

repels males

2-sec-Butyl-4,5-dihydrothiazole Mouse Attracts females and VNO

repels males; induces

2,3-Dehydro-exobrevicomin Mouse Attracts females and VNO

repels males; induces

2-Methylbut-2-enal Rabbit Attracts pups MOE

to nipples

Dimethyl disulfide Hamster Induces copulation VNO

in males

(Z)-7-Dodecenyl acetate Asian elephant Attracts males ?

5-a-Androst-16-en-3-one Pig Facilitates mating MOE

in females

Sodefrin Newt Attracts females VNO

apical

basal

Key signaling molecules identified in VNO neurons

Apical neurons:

V1R, Gi2, TRPC2

Basal neurons:

V2R, Go, TRPC2

V1R/V2R Pheromone

Transduction cascade in VNO neurons

a

bg

PIP2

IP3

DAGTRPC2

Ca2+

AA

AA-gated channel

CaM

DAG lipaseDAG

PLC

Gi2/G0

Species V1R V2R TRPC2

Man 5 (115) 0 (20) 0 (1)

Chimpanzee 0 (116) 0 (17) 0 (1)

Macaque ? 0 (11) ?

Mouse 187 (121) 121 (158) 1

Rat 106 (66) 79 (142) 1

Dog 8 (33) 0 (9) 1

Caw 40 (45) 0 (16) ?

Opossum 98 (30) 86 (79) ?

Number of genes (pseudogenes)

Volatile male

gender signal

Do chemical communications between humans exist?

After Spehr et al., Cell. Mol. Life Sci., 2006

Common odorants

Electronic nose

Electronic nose applications

I. Medicine

1. Biological liquids analysis

2. Disease detection

II. Security

1. Detection of individuals

2. Detection of explosives and narcotics

3. Detection of poisons and toxins in air and water

4. Detection of bacterial contaminations

III. Crime Investigation

1. Detection of individuals

2. Detection of poisons and toxins in a victom body

3. Detection of bacterial contaminations

III. Food Industry

1. Analysis of food and drink quality

2. Detection of hazardous components

IV. Perfumery industry

1. Perfume quality control

2. New odor compositions

Odor detection

Sensory

information

processing

Odor

recognition

Olfactory sensory

neurons

Olfactory bulb Olfactory cortex

Decision

making

Brain

Odor

Information processing in olfactory system

Schematic diagram of an electronic nose

Sensor 1

Data

processing

pattern

recognition

**************

Odor Sensor 2

Sensor NData base

system

Decision

making

Surface plasmon resonance

Input lightTransmitted

light

metal layer

sensing layer plasmon resonance

glass fiber

cladding

SPR sensor

odorant

Conductivity sensor

Sensing layer

Metal electrode

Conductance gauge

Substrate

odorant

Piezoelectric sensor

Quartz disc

electrode

Polymer coatingodorant

Salmonella typhimurium

Streppneumonia

Shigellaflexneri

Salmonellaenteritidis

Hemophilusinfluenza

Enterococcusfaecalis

E. Coli O157E. Coli

Fresh Nutrient

Staph aureus Pseudomonasaeruginosa

Klebsiellapneumonia

VaporPrint analysis of Infectious Bacteria Cultures

Electric Sensor Technology

Salmonella typhimurium

Fresh Nutrient

Спасибо за внимание!