Evaluation of an independent prestin mouse model derived from the 129S1 strain.
Studies using the prestin knockout mouse indicate that removal of the outer hair cell (OHC) motor protein is associated with loss of sensitivity, frequency selectivity and somatic electromotility. Here we provide data obtained from another prestin mouse model that was produced commercially. In vivo electrical recordings from the round window indicate that the phenotype is similar to that of the original knockout generated by the Zuo group at St. Jude Children's Research Hospital. Hence, compound action potential (CAP) thresholds are shifted in a frequency-dependent manner and CAP tuning curves at 12 kHz are flat for masker frequencies between 3 and 18 kHz. Although CAP input-output functions at 6 kHz show a shift in sensitivity at low levels, responses approach wild-type magnitudes at high levels where the cochlear amplifier has less influence. In order to confirm that the loss of sensitivity and frequency selectivity is due to loss of prestin, we performed immunohistochemistry using a prestin antibody. Cochlear segments from homozygous mutant mice showed no fluorescence, while wild-type mice displayed a fluorescent signal targeted to the OHC's lateral membrane. Absence of prestin protein was confirmed using LDS-PAGE/Western blot analysis. These results indicate that the loss of function phenotype is associated with loss of prestin protein. Lack of prestin protein also results in a shortening of OHC length to approximately 60% of wild-type, similar to that reported previously by Liberman's group. The linkage shown between the loss of prestin protein and abnormal cochlear function validates the original knockout and attests to the importance of OHC motor function in the auditory periphery.
Department of Communication Sciences and Disorders, The Hugh Knowles Center, Evanston, IL, USA. m-cheatham@northwestern.edu
M A
MA
Cheatham
J
J
Zheng
K H
KH
Huynh
G G
GG
Du
R M
RM
Edge
C T
CT
Anderson
J
J
Zuo
A F
AF
Ryan
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Dallos
D000818:Animals; D001309:Auditory Threshold / Q000502:physiology; D003055:Cochlear Microphonic Potentials / Q000235:genetics; D004195:Disease Models, Animal; D005091:Exons / Q000235:genetics; D018390:Gene Targeting; D005838:Genotype; D018072:Hair Cells, Auditory, Outer / Q000473:pathology / Q000502:physiology*; D006579:Heterozygote; D006720:Homozygote; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008817:Mice, Mutant Strains; D008822:Mice, Transgenic; D018613:Microscopy, Confocal; D020409:Molecular Motor Proteins / Q000235:genetics*; D010641:Phenotype; D012333:RNA, Messenger / Q000235:genetics
D020409:Molecular Motor Proteins; C498720:Pres protein, mouse; D012333:RNA, Messenger