Colony-Wide Analysis of Mouse Auditory Brainstem Responses (II): Maturational, Gender and Aging Effects in C57Bl/6J and CBA/CaJ MicePaul Allen1, John Housel2, Jason Moore1, William O'Neill1,2, Peter Rivoli1, James Ison1

1Department of Brain & Cognitive Sciences, University of Rochester;2 Dept of Neurobiology & Anatomy, University of Rochester School of Medicine and Dentistry; Rochester, NYARO 2005

433

Introduction

CBA/CaJ: (710 recordings, 388 from Males, 322 from Females)

C57Bl/6: (484 recordings, 244 from Males, 240 from Females)

Auditory brainstem response (ABR)

Routinely measured in conjunction with behavioral, electrophysiological or histological experiments

For methods see presentation 437

Multiple ABRs were accepted for an animal only if there was a minimum of 60 days between recordings

Animals undergoing drug treatment etc. are excluded

Superthreshold ABR is assessed via signal to noise

S/N ratio calculated from the RMS of 2kHz high-pass filtered waveforms

RMS windows: Pre-response = 0.75-1.5 ms, Evoked Response = 1.5-10 ms

S/N calculated for a subset of mice (using TDT System II apparatus)

CBA/CaJ (N=259, 145 Males and 114 Females)

C57Bl/6 (N=99, 39 Males and 60 Females) Supported by NIA Grant #AG09524 and The Schmitt Program on Integrative Brain Research

Subjects and Methods

CBA/CaJ

3 6 12 24 32 480

10

20

30

40

50

60

70

80

90

15-22 mo23-27 mo28-34 mo

7-14 mo

1 mo

SEM

Frequency (Hz)

Leve

l (dB

SP

L)

Age distribution of CBA mice

0 4 8 12 16 20 24 28 32 360

25

50

75

100

125

150

175

Female

Male

Age (mo)

N

Age distribution of C57 mice

0 1 2 3 4 5 6 7 8 9 10 11 120

20

40

60

80

100

120

Female

Male

Age (mo)

N

1 2 3 4 5-6 7-14 15-22 23-27 28-34-0.6

-0.5

-0.4

-0.3

-0.2

-0.1

-0.0

0.1

0.2

0.3

0.4

0.5

0.6 3 kHz

12 kHz24 kHz32 kHz48 kHz

6 kHz

Male Advantage

Female Advantage

SEM

Age (mo)

AB

R R

MS

Rat

io

1 2 3 5 7 8 10 120

10

20

30

40

50

60

70

80

90

100

6 kHz Male

3 kHz Female

48 kHz Female48 kHz Male

3 kHz Male

6 kHz Female

Age (Months)

AB

R t

hres

hold

(dB

SP

L)

SEM

C57Bl/6J

3 6 12 24 32 480

10

20

30

40

50

60

70

80

90

100

1 mo

3 mo

6 mo8 mo

12 moSEM

Frequency (Hz)

Leve

l (dB

SP

L)

1 2 3 5 7 8 10 12

12 kHz Female

16 kHz Female16 kHz Male

32 kHz Female

24 kHz Female24 kHz Male

32 kHz Male

0

10

20

30

40

50

60

70

80

90

100

12 kHz Male

Age (Months)

AB

R t

hres

hold

(dB

SP

L)

SEM

1 2 3 4 5-6 7-14 15-22 23-27 28-340

10

20

30

40

50

60

70

80

12 kHz Male12 kHz Female

32 kHz Female32 kHz Male

24 kHz Male24 kHz Female

Age (Months)

AB

R t

hres

hold

(dB

SP

L)

SEM

1 2 3 4 5-6 7-14 15-22 23-27 28-340

10

20

30

40

50

60

70

80

90

3 kHz Female3 kHz Male

6 kHz Female

48 kHz Female48 kHz Male

6 kHz Male

Age (Months)

AB

R t

hres

hold

(dB

SP

L)

SEM

C5712 kHz, 90 dB SPL

1 2 3 4 5 6 7 8-3.0×10-06

-2.0×10-06

-1.0×10-06

-2.3×10-13

1.0×10-06

2.0×10-06

3.0×10-06

4.0×10-06

5.0×10-06

6.0×10-06

7.0×10-06

Early: 2-4 moMiddle: 4-6 moLate: 6-8 mo

Time (ms)

Vo

ltag

e

2-4 mo

1 2 3 4 5 6 7 8

-2.0×10 -6

0

2.0×10 -6

4.0×10 -6

6.0×10 -6 MaleFemale

4-6 mo

1 2 3 4 5 6 7 8

-2.0×10 -6

0

2.0×10 -6

4.0×10 -6

6.0×10 -6

MaleFemale

6-8 mo

1 2 3 4 5 6 7 8

-2.0×10 -6

0

2.0×10 -6

4.0×10 -6

6.0×10 -6 MaleFemale

12 kHz, 90 dB SPLCBA12 kHz 90 dB SPL

1 2 3 4 5 6 7 8-4.0×10-6

-2.0×10-6

0

2.0×10-6

4.0×10-6

6.0×10-6

Young: 2-4 moMiddle-aged: 11-13 moOld: 23-25 mo

Time (msec)

AB

R (

V)

0

2-4 mo

1 2 3 4 5 6 7 8

-4.5×10 -6

-3.0×10 -6

-1.5×10 -6

1.5×10 -6

3.0×10 -6

4.5×10 -6

6.0×10 -6

7.5×10 -6

0

MaleFemale

0

11-13 mo

1 2 3 4 5 6 7 8

-4.5×10 -6

-3.0×10 -6

-1.5×10 -6

1.5×10 -6

3.0×10 -6

4.5×10 -6

6.0×10 -6

7.5×10 -6

0

MaleFemale

0

23-25 mo

1 2 3 4 5 6 7 8

-4.5×10 -6

-3.0×10 -6

-1.5×10 -6

1.5×10 -6

3.0×10 -6

4.5×10 -6

6.0×10 -6

7.5×10 -6

0

MaleFemale

Changes in ABR Waveforms with Age Changes in ABR Waveforms with Hearing-loss

0 1 2 3 4 5 6 7 8 90

1

2

3

4

5

6

7

83 kHz

12 kHz24 kHz32 kHz48 kHz

6 kHz

Age (mo)

AB

R R

MS

Rat

io

SEM

2 3 4 5 6 7 8

-0.6

-0.5

-0.4

-0.3

-0.2

-0.1

-0.0

0.1

0.2

0.3

0.4

0.5

0.6

3 kHz

12 kHz24 kHz32 kHz48 kHz

6 kHz

Female Advantage

Male Advantage

SEM

Age (mo)

AB

R R

MS

Rat

io d

iffe

ren

ce

1 2 3 4 5-6 7-14 15-22 23-27 28-34

1

2

3

4

5

63 kHz

12 kHz24 kHz32 kHz48 kHz

6 kHzSEM

Age (mo)

AB

R R

MS

Rat

io

0 1 2 3 4 5 6 7 8 9 10

-5.0×10 -6

-2.5×10 -6

0

2.5×10 -6

5.0×10 -6

Pre-Response RMS: 4.4125e-07Response RMS: 3.1721e-08Ratio: 13.9103

rawfiltered

Time (ms)

AB

R (

V)

22 M - 17 F41 M - 22 F9 M - 9 F27 M - 21 F11 M - 10 F35 M - 35 F

Leve

l (dB

SP

L)Le

vel (

dB S

PL)

2 mo 5-7 mo 8-14 mo 15-22 mo 23-27 mo 28-34 mo

16 M - 20 F 19 M -17 F 1 M - 8 F 3 M - 7 F 0 M - 8 F

Leve

l (dB

SP

L)Le

vel (

dB S

PL)

2 mo 3 mo 5 mo 7 mo 8 mo

ABR Thresholds from 1 to 30 Months of Age ABR Thresholds: 1 to 12 Months of Age

ABR Ratio Plots

Difference in Ratio

Male - Female

ABR Ratio Plots

Difference in Ratio

Male - Female

Changes in Superthreshold ABR with Hearing-lossAging-related Changes in Superthreshold ABR

ABR Ratio vs Age

90dB SPL

Difference in Ratio

Male - Female

ABR Ratio vs Age

90dB SPL

Difference in Ratio

Male - Female

CBA/CaJ and C57BL/6J mice provide useful models for contrasting rapid degenerative vs. slow age-related hearing loss. The descriptions of ABR hearing measures in these mice are mostly obtained in small groups (save for Li and Borg, 1991), and concerned with thresholds (save for Hunter and Willott, 1987). Our colony at the University of Rochester provides these mice to investigators focused on age-related changes in complex auditory processing. Both threshold and superthreshold ABR measures are taken on many mice at ages that meet the special needs of particular protocols. Here we describe ABR measures collected over 6 years, in a total of 570 CBA mice (267F, 303M) and 353 C57 (161F, 192M), most born in the vivarium from Jackson Lab breeding stock. The youngest mice for which records are available were 27 days of age while the oldest mice were 1046 days of age for the CBA (n = 2) and 787 days of age for the C57 (n=2). The data we present are grouped in age bins providing sufficiently large samples of males and females to minimize potential cohort and procedural differences across time.

• Reduction in P1 amplitudes (as seen for 12kHz, 90dB SPL)

• Lengthening of Inter-peak intervals

(Note that wave forms are aligned on P1)

• Subtle changes in ABR waveform with age

Change in appearance of P1a/N1 complex

• No sex effect on ABR waveforms

• No threshold loss (and possible maturational gain?) up to 14 mo of age

• Progressive loss, equal across frequency, beyond 14 mo of age

• No systematic sex differences

• Superthreshold S/N ratio plots show substantial decline with age in first year of life, despite negligible threshold changes

Central neural aging?

Artifact of changes in head size, or dielectric properties of cranium?

• Possible weak female advantage seen in S/N color plots up to 22 months

• S/N ratios for 90 dB highlight the progressive decline into old age and lack of systematic sex differences with age and frequency

Left: “Classic” frequency across age Right: Age effect across frequency 1) Initial slow loss for all frequencies, then delayed abrupt loss, higher first2) Old females are slightly more sensitive to low frequencies

Reduction in P1 amplitude (shown for 12kHz, 90 dB)(Note that waveforms have been aligned on P1)

ABR waveforms change with hearing-loss

Inter-peak intervals remain stable (except P2, P4)

Females have slightly larger ABR peaks (Head size?)

Superthreshold S/N ratios show progressive high frequency hearing loss in parallel with the elevated ABR thresholds. The 90 dB S/N data show a progressive decline with age for all frequencies

Possible female advantage is seen in S/N ratios above threshold, and is consistent with threshold data

The initial age-related gradual hearing loss across frequencies is consistent with, e.g., Hequembourg & Liberman, (2001), but the data favoring female over male inconsistent with Willott & Bross (2004) or Henry (2002).