The long-term objectives are to understand the mechanisms of mechanotransduction in auditory hair cells and delineate the factors underlying the cochlea's tonotopic organization. Experiments will focus on the attributes and molecular composition of the mechanotransducer (MT) channel and its modulation by Ca2+. Hair cell responses will be measured in the isolated cochleas of both mouse and chicken. A feature of the work is the use of mouse mutants combined with transfection of cultured auditory epithelia.
Specific aims are: (1) to record MT currents in cochlear hair cells from mice with mutations in the transmembrane channel-like (Tmc) proteins Tmc1 and Tmc2, which are molecular candidates for the MT channel. Available mouse mutants will be studied as well as those with engineered point mutations which will be transfected into cochlear cultures;(2) to examine the Ca2+ dependence and mechanism of MT channel adaptation in mammalian hair cells, testing the hypothesis that it differs from that established in non-mammals. Ca2+ will be manipulated by internal perfusion and photolysis of caged Ca2+. Mice with mutations in myosin VIIa will also be characterized to determine the contribution of this myosin to adaptation;(3) to define the function of the hair cell Ca2+-binding protein oncomodulin in Ca2+ buffering and development of hair cells by generating conditional mutants and exploring their properties;the hypothesis is that embryonic expression of oncomodulin is essential for hair cell development but that post-natal loss is without effect;(4) to record MT currents and measure hair bundle mechanics of short (outer) hair cells in the chick auditory papilla as a likely but unproven site where active hair bundle motion is used to augment frequency selectivity. The interaction between active bundle motion derived from channel gating and a possible prestin-induced motility will be used to assess the contributions of the two processes to amplification and frequency tuning. Comparison with the properties of the mammalian outer hair cells will provide insight into the evolution of cochlear amplification in amniotes;(5) to examine the embryonic maturation of frequency tuning in the chicken auditory papilla, testing a previously reported shift in best frequency;also charting the embryonic development of the MT current, the role of Tmc proteins and of parvalbumin-3, the avian equivalent of oncomodulin. It is hoped that the results will supply evidence on the molecular composition of the hair cell transduction apparatus and will yield information about proteins that are mutated in certain forms of human genetic deafness.

Public Health Relevance

Severe to profound hearing loss, largely attributable to injury to the sensory hair cells, affects 1 in 1,000 newborns, and 60% of people older than 70 years, and has multiple causes, genetic, environmentally-induced and age-related. The basic mechanisms of hair cell damage and death are in most cases not well understood and the work will address these mechanisms in two ways. Firstly by studying mouse mutants that have human equivalents such as Tmc1, which, with over 30 mutations, is one of the most common causes of genetic hearing loss;and myosin VIIa, the most prevalent type of Usher type 1 syndrome. Secondly, by investigating intracellular Ca2+ regulation, defects in which may underlie hair cell death due to ototoxic drugs and aging.

Agency
National Institute of Health (NIH)
Institute
National Institute on Deafness and Other Communication Disorders (NIDCD)
Type
Research Project (R01)
Project #
2R01DC001362-23
Application #
8575785
Study Section
Auditory System Study Section (AUD)
Program Officer
Cyr, Janet
Project Start
1992-01-01
Project End
2018-12-31
Budget Start
2014-01-01
Budget End
2014-12-31
Support Year
23
Fiscal Year
2014
Total Cost
$450,460
Indirect Cost
$139,457
Name
University of Wisconsin Madison
Department
Neurosciences
Type
Schools of Medicine
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
Fettiplace, Robert; Kim, Kyunghee X (2014) The physiology of mechanoelectrical transduction channels in hearing. Physiol Rev 94:951-86
Beurg, Maryline; Kim, Kyunghee X; Fettiplace, Robert (2014) Conductance and block of hair-cell mechanotransducer channels in transmembrane channel-like protein mutants. J Gen Physiol 144:55-69
Kim, Kyunghee X; Beurg, Maryline; Hackney, Carole M et al. (2013) The role of transmembrane channel-like proteins in the operation of hair cell mechanotransducer channels. J Gen Physiol 142:493-505
Tan, Xiaodong; Beurg, Maryline; Hackney, Carole et al. (2013) Electrical tuning and transduction in short hair cells of the chicken auditory papilla. J Neurophysiol 109:2007-20
Kim, Kyunghee X; Fettiplace, Robert (2013) Developmental changes in the cochlear hair cell mechanotransducer channel and their regulation by transmembrane channel-like proteins. J Gen Physiol 141:141-8
Beurg, Maryline; Tan, Xiaodong; Fettiplace, Robert (2013) A prestin motor in chicken auditory hair cells: active force generation in a nonmammalian species. Neuron 79:69-81
Johnson, Stuart L; Beurg, Maryline; Marcotti, Walter et al. (2011) Prestin-driven cochlear amplification is not limited by the outer hair cell membrane time constant. Neuron 70:1143-54
Mahendrasingam, Shanthini; Beurg, Maryline; Fettiplace, Robert et al. (2010) The ultrastructural distribution of prestin in outer hair cells: a post-embedding immunogold investigation of low-frequency and high-frequency regions of the rat cochlea. Eur J Neurosci 31:1595-605
Nam, Jong-Hoon; Fettiplace, Robert (2010) Force transmission in the organ of Corti micromachine. Biophys J 98:2813-21
Beurg, Maryline; Nam, Jong-Hoon; Chen, Qingguo et al. (2010) Calcium balance and mechanotransduction in rat cochlear hair cells. J Neurophysiol 104:18-34

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