A major first step in audition is the conversion of mechanical deflection of the sensory hair bundle into an electrical signal that drives the hair cell receptor potential that in turn controls cochlear amplification (outer hair cells) and synaptic transmission (inner hair cells). Alterations in any step of this process will degrade signal processing throughout the auditory pathway. During the previous funding period, we demonstrated that mammalian cochlea hair bundles do not move coherently, meaning that there were both temporal and magnitude differences between stereocilia motion depending on the mode of stimulation. The lack of coherence leads to very different mechanotransducer current responses, which are predicted to generate very different receptor potentials. This finding drives a major component of the present proposal which is focused upon identifying how hair bundles move in situ, to better understand the properties of the physiologically driven mechanotransducer currents. We have developed new technology including imaging bundle movement in situ at high speeds as well as image processing algorithms that can detect motions at below 2 nm. We will use these new technologies as well as high speed calcium imaging, new fluid jet and stiff probe stimulating devices and electrophysiological tools to investigate stereocilia motion. Results from these experiments will shed light onto why outer hair cells are embedded in the tectorial membrane and inner hair cells are not. We will also identify the hair bundle and tectorial membrane mechanical properties responsible for regulating hair bundle motion. During the past funding period we also identified several mechanisms by which the lipid membrane either directly or indirectly alters mechanotransduction currents. We have developed new technologies including fluorescence recovery after photobleaching to directly assess the mechanical properties of the lipid bilayer as a means of identifying the underlying mechanisms for regulating the mechanotransducer channel. We will use these new technologies coupled with pharmacological manipulations of the lipid bilayer to investigate the role of specific lipids and their mechanical properties in modulating hair cell MET.

Public Health Relevance

The hair bundle of sensory hair cells is the site where sounds vibrations is converted into an electrical signal. Hair bundle proteins underlie many genetically driven hearing loss pathologies. Similarly, the hair bundle is a target for noise induced as well as ageing induced hearing loss and so identifying the mechanisms by which the hair bundle works normally is important to develop strategies for damage prevention and system repair. We have demonstrated that the hair cell output is directly dependent on the mode of hair bundle stimulation because the stereocilia within each hair bundle are not tightly coupled mechanically. During this funding period, we will determine how hair bundles normally move and what hair bundle and tectorial membrane properties drive the motion. We are also investigating the role of the lipid bilayer in modulating mechanotransduction. Although the lipid bilayer is known to modulate ion channel function in many systems, either by directly affecting channel activity or indirectly modifying behavior through its mechanical properties, investigations of the hair cell stereocilia lipid environment are in their infancy. This proposal will explore the roles of PIP2, cholesterol, sphingomyelin and conical lipids on membrane fluidity and hair bundle function.

Agency
National Institute of Health (NIH)
Institute
National Institute on Deafness and Other Communication Disorders (NIDCD)
Type
Research Project (R01)
Project #
5R01DC003896-24
Application #
10075251
Study Section
Auditory System Study Section (AUD)
Program Officer
Cyr, Janet
Project Start
1999-01-18
Project End
2022-12-31
Budget Start
2021-01-01
Budget End
2021-12-31
Support Year
24
Fiscal Year
2021
Total Cost
Indirect Cost
Name
Stanford University
Department
Otolaryngology
Type
Schools of Medicine
DUNS #
009214214
City
Stanford
State
CA
Country
United States
Zip Code
94305
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Larsen, T; Doll, J C; Loizeau, F et al. (2017) Rise Time Reduction of Thermal Actuators Operated in Air and Water through Optimized Pre-Shaped Open-Loop Driving. J Micromech Microeng 27:
Kazmierczak, Marcin; Kazmierczak, Piotr; Peng, Anthony W et al. (2017) Pejvakin, a Candidate Stereociliary Rootlet Protein, Regulates Hair Cell Function in a Cell-Autonomous Manner. J Neurosci 37:3447-3464
Peng, Anthony W; Gnanasambandam, Radhakrishnan; Sachs, Frederick et al. (2016) Adaptation Independent Modulation of Auditory Hair Cell Mechanotransduction Channel Open Probability Implicates a Role for the Lipid Bilayer. J Neurosci 36:2945-56
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Sundaresan, Srividya; Kong, Jee-Hyun; Fang, Qing et al. (2016) Thyroid hormone is required for pruning, functioning and long-term maintenance of afferent inner hair cell synapses. Eur J Neurosci 43:148-61
Peng, Anthony W; Ricci, Anthony J (2016) Glass Probe Stimulation of Hair Cell Stereocilia. Methods Mol Biol 1427:487-500
Soons, Joris A M; Ricci, Anthony J; Steele, Charles R et al. (2015) Cytoarchitecture of the mouse organ of corti from base to apex, determined using in situ two-photon imaging. J Assoc Res Otolaryngol 16:47-66
Alyono, Jennifer C; Corrales, C Eduardo; Huth, Markus E et al. (2015) Development and characterization of chemical cochleostomy in the Guinea pig. Otolaryngol Head Neck Surg 152:1113-8
Song, Yohan; Xia, Anping; Lee, Hee Yoon et al. (2015) Activity-dependent regulation of prestin expression in mouse outer hair cells. J Neurophysiol 113:3531-42

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