Hearing loss affects approximately 25 Million Americans. It also affects approximately 35% of those 65 and older. The primary cause of hearing loss in this group is a result of a loss of hair cells. Our interest is in understanding the molecular events that determine hair cell function, and how these events are controlled. We anticipate that such an understanding will allow rational therapies to be developed to treat hair cell loss. Hair cells are the primary transducers of sound serving to convert the mechanical energy of sound to a coded a neuronal one. In this proposal we are seeking to understand the molecular basis of the large conductance potassium channel as it affects hair cell function. Specifically, we are trying to elucidate the molecular determinants of how these channels bring about electrical tuning, a mechanism of frequency discrimination. We are also interested in determining the molecular basis of how they are clustered and co-localized with the voltage gated Ca channel at the basolateral surface of hair cells. The loss of the large conductance potassium channel in mice results in progressive hearing loss. We propose to extend previous work that has identified variations in the primary structure of the protein, and now seek to determine how interacting proteins influence its function. We have already isolated several proteins from the cochlea that interact with a limited portion of this channel using the yeast 2 hybrid technique. We will now determine how these interacting proteins affect the kinetic properties of this channel. We will also determine how these proteins affect it basolateral sorting, clustering and co-localization with voltage gated Ca channels. In addition, we will extend our successful yeast 2 hybrid approach to identify other proteins that interact with entire BK channel. These proteins too will be subject to similar assays. For these experiments we wilt use the chick as a model and will use a modular set of assays that we have developed to test their significance in affecting these physiological processes.

Agency
National Institute of Health (NIH)
Institute
National Institute on Deafness and Other Communication Disorders (NIDCD)
Type
Research Project (R01)
Project #
5R01DC007894-04
Application #
7628038
Study Section
Auditory System Study Section (AUD)
Program Officer
Cyr, Janet
Project Start
2006-07-01
Project End
2011-06-30
Budget Start
2009-07-01
Budget End
2010-06-30
Support Year
4
Fiscal Year
2009
Total Cost
$356,876
Indirect Cost
Name
Yale University
Department
Neurology
Type
Schools of Medicine
DUNS #
043207562
City
New Haven
State
CT
Country
United States
Zip Code
06520
Fleming, Matthew R; Brown, Maile R; Kronengold, Jack et al. (2016) Stimulation of Slack K(+) Channels Alters Mass at the Plasma Membrane by Triggering Dissociation of a Phosphatase-Regulatory Complex. Cell Rep 16:2281-8
Zhang, Yalan; Zhang, Xiao-Feng; Fleming, Matthew R et al. (2016) Kv3.3 Channels Bind Hax-1 and Arp2/3 to Assemble a Stable Local Actin Network that Regulates Channel Gating. Cell 165:434-448
Surguchev, Alexei; Bai, Jun-Ping; Joshi, Powrnima et al. (2012) Hair cell BK channels interact with RACK1, and PKC increases its expression on the cell surface by indirect phosphorylation. Am J Physiol Cell Physiol 303:C143-50
Bai, Jun-Ping; Surguchev, Alexei; Joshi, Powrnima et al. (2012) CDK5 interacts with Slo and affects its surface expression and kinetics through direct phosphorylation. Am J Physiol Cell Physiol 302:C766-80
Bai, Jun-Ping; Surguchev, Alexei; Navaratnam, Dhasakumar (2011) ýý4-subunit increases Slo responsiveness to physiological Ca2+ concentrations and together with ýý1 reduces surface expression of Slo in hair cells. Am J Physiol Cell Physiol 300:C435-46
Bian, Shumin; Bai, Jun-Ping; Chapin, Hannah et al. (2011) Interactions between ?-catenin and the HSlo potassium channel regulates HSlo surface expression. PLoS One 6:e28264
Frucht, Corey S; Uduman, Mohamed; Kleinstein, Steven H et al. (2011) Gene expression gradients along the tonotopic axis of the chicken auditory epithelium. J Assoc Res Otolaryngol 12:423-35
Frucht, Corey S; Santos-Sacchi, Joseph; Navaratnam, Dhasakumar S (2011) MicroRNA181a plays a key role in hair cell regeneration in the avian auditory epithelium. Neurosci Lett 493:44-8
Bai, Jun-Ping; Surguchev, Alexei; Bian, Shumin et al. (2010) Combinatorial cysteine mutagenesis reveals a critical intramonomer role for cysteines in prestin voltage sensing. Biophys J 99:85-94
Bian, Shumin; Koo, Bon W; Kelleher, Stephen et al. (2010) A highly expressing Tet-inducible cell line recapitulates in situ developmental changes in prestin's Boltzmann characteristics and reveals early maturational events. Am J Physiol Cell Physiol 299:C828-35

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