17 results on '"Frolenkov, Gregory I."'
Search Results
2. Dynamic Assembly of Surface Structures in Living Cells
- Author
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Gorelik, Julia, Shevchuk, Andrew I., Frolenkov, Gregory I., Diakonov, Ivan A., Lab, Max J., Kros, Corné J., Richardson, Guy P., Vodyanoy, Igor, Klenerman, David, and Korchev, Yuri E.
- Published
- 2003
3. γ-Actin Is Required for Cytoskeletal Maintenance but Not Development
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Belyantseva, Inna A., Perrin, Benjamin J., Sonnemann, Kevin J., Zhu, Mei, Stepanyan, Ruben, McGee, JoAnn, Frolenkov, Gregory I., Walsh, Edward J., Friderici, Karen H., Friedman, Thomas B., Ervasti, James M., and Frieden, Carl
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- 2009
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4. Unbalanced bidirectional radial stiffness gradients within the organ of Corti promoted by TRIOBP.
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Babahosseini, Hesam, Belyantseva, Inna A., Yousaf, Rizwan, Tona, Risa, Hadi, Shadan, Inagaki, Sayaka, Wilson, Elizabeth, Shin-ichiro Kitajiri, Frolenkov, Gregory I., Friedman, Thomas B., and Cartagena-Rivera, Alexander X.
- Subjects
CORTI'S organ ,HAIR cells ,ATOMIC force microscopy ,CELLULAR mechanics ,FOCUSED ion beams - Abstract
Hearing depends on intricate morphologies and mechanical properties of diverse inner ear cell types. The individual contributions of various inner ear cell types into mechanical properties of the organ of Corti and the mechanisms of their integration are yet largely unknown. Using sub-100-nm spatial resolution atomic force microscopy (AFM), we mapped the Young's modulus (stiffness) of the apical surface of the different cells of the freshly dissected P5-P6 cochlear epithelium from wild-type and mice lacking either Trio and F-actin binding protein (TRIOBP) isoforms 4 and 5 or isoform 5 only. Variants of TRIOBP are associated with deafness in human and in Triobp mutant mouse models. Remarkably, nanoscale AFM mapping revealed unrecognized bidirectional radial stiffness gradients of different magnitudes and opposite orientations between rows of wild-type supporting cells and sensory hair cells. Moreover, the observed bidirectional radial stiffness gradients are unbalanced, with sensory cells being stiffer overall compared to neighboring supporting cells. Deafness-associated TRIOBP deficiencies significantly disrupted the magnitude and orientation of these bidirectional radial stiffness gradients. In addition, serial sectioning with focused ion beam and backscatter scanning electron microscopy shows that a TRIOBP deficiency results in ultrastructural changes of supporting cell apical phalangeal microfilaments and bundled cortical F-actin of hair cell cuticular plates, correlating with messenger RNA and protein expression levels and AFM stiffness measurements that exposed a softening of the apical surface of the sensory epithelium in mutant mice. Altogether, this additional complexity in the mechanical properties of the sensory epithelium is hypothesized to be an essential contributor to frequency selectivity and sensitivity of mammalian hearing. [ABSTRACT FROM AUTHOR]
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- 2022
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5. Myosin-XVa Controls Both Staircase Architecture and Diameter Gradation of Stereocilia Rows in the Auditory Hair Cell Bundles.
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Hadi, Shadan, Alexander, Andrew J., Vélez-Ortega, A. Catalina, and Frolenkov, Gregory I.
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HAIR cells ,FOCUSED ion beams ,STAIRCASES ,DIAMETER ,SCANNING electron microscopy - Abstract
Mammalian hair cells develop their mechanosensory bundles through consecutive phases of stereocilia elongation, thickening, and retraction of supernumerary stereocilia. Many molecules involved in stereocilia elongation have been identified, including myosin-XVa. Significantly less is known about molecular mechanisms of stereocilia thickening and retraction. Here, we used scanning electron microscopy (SEM) to quantify postnatal changes in number and diameters of the auditory hair cell stereocilia in shaker-2 mice (Myo15sh2) that lack both "long" and "short" isoforms of myosin-XVa, and in mice lacking only the "long" myosin-XVa isoform (Myo15∆N). Previously, we observed large mechanotransduction current in young postnatal inner (IHC) and outer (OHC) hair cells of both these strains. Stereocilia counts showed nearly identical developmental retraction of supernumerary stereocilia in control heterozygous, Myo15sh2/sh2, and Myo15∆N/∆N mice, suggesting that this retraction is largely unaffected by myosin-XVa deficiency. However, myosin-XVa deficiency does affect stereocilia diameters. In control, the first (tallest) and second row stereocilia grow in diameter simultaneously. However, the third row stereocilia in IHCs grow only until postnatal day 1-2 and then become thinner. In OHCs, they also grow slower than taller stereocilia, forming a stereocilia diameter gradation within a hair bundle. The sh2 mutation disrupts this gradation and makes all stereocilia nearly identical in thickness in both IHCs and OHCs, with only subtle residual diameter differences. All Myo15sh2/sh2 stereocilia grow postnatally including the third row, which is not a part of normal development. Serial sections with focused ion beam (FIB)-SEM confirmed that diameter changes of Myo15sh2/sh2 IHC and OHC stereocilia resulted from corresponding changes of their actin cores. In contrast to Myo15sh2/sh2, Myo15∆N/∆N hair cells develop prominent stereocilia diameter gradation. Thus, besides building the staircase, the short isoform of myosin-XVa is essential for controlling the diameter of the third row stereocilia and formation of the stereocilia diameter gradation in a hair bundle. [ABSTRACT FROM AUTHOR]
- Published
- 2020
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6. Building and repairing the stereocilia cytoskeleton in mammalian auditory hair cells.
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Vélez-Ortega, A. Catalina and Frolenkov, Gregory I.
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HAIR cells , *BUILDING repair , *CYTOSKELETON , *LIFE spans , *COCHLEA - Abstract
Abstract Despite all recent achievements in identification of the molecules that are essential for the structure and mechanosensory function of stereocilia bundles in the auditory hair cells of mammalian species, we still have only a rudimentary understanding of the mechanisms of stereocilia formation, maintenance, and repair. Important molecular differences distinguishing mammalian auditory hair cells from hair cells of other types and species have been recently revealed. In addition, we are beginning to solve the puzzle of the apparent life-long stability of the stereocilia bundles in these cells. New data link the stability of the cytoskeleton in the mammalian auditory stereocilia with the normal activity of mechanotransduction channels. These data suggest new ideas on how a terminally-differentiated non-regenerating hair cell in the mammalian cochlea may repair and tune its stereocilia bundle throughout the life span of the organism. Highlights • Differentially expressed proteins regulate the hair bundle staircase architecture. • The stability of the stereocilia actin core depends on intrastereocilia Ca2+ levels. • Changes in intrastereocilia Ca2+ levels may trigger signals for stereocilia repair. • Ca2+-sensitive proteins within stereocilia could be potential therapeutic targets. [ABSTRACT FROM AUTHOR]
- Published
- 2019
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7. Molecular Remodeling of Tip Links Underlies Mechanosensory Regeneration in Auditory Hair Cells.
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Indzhykulian, Artur A., Stepanyan, Ruben, Nelina, Anastasiia, Spinelli, Kateri J., Ahmed, Zubair M., Belyantseva, Inna A., Friedman, Thomas B., Barr-Gillespie, Peter G., and Frolenkov, Gregory I.
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HAIR cells ,CORTI'S organ ,ACETIC acid ,FATTY acids ,GENETIC transduction - Abstract
Backscatter scanning electron microscopy and conventional whole cell patch-clamp experiments reveal a two-step mechanism for the regeneration of tip links, the crucial element of mechanotransduction machinery in the hair cells of the inner ear. [ABSTRACT FROM AUTHOR]
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- 2013
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8. Fast Adaptation and Ca2+ Sensitivity of theMechanotransducer Require Myosin-XVa in Inner But Not Outer Cochlear Hair Cells.
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Stepanyan, Ruben and Frolenkov, Gregory I.
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NEUROBIOLOGY , *TRANSDUCERS , *CHANNELS (Structural members) , *HAIR cells , *COCHLEA , *MYOSIN , *LABORATORY mice - Abstract
In inner ear hair cells, activation of mechanotransduction channels is followed by extremely rapid deactivation that depends on the influx of Ca2+ through these channels. Although the molecular mechanisms of this "fast" adaptation are largely unknown, the predominant models assume Ca22+ sensitivity as an intrinsic property of yet unidentified mechanotransduction channels. Here, we examined mechanotransduction in the hair cells of young postnatal shaker 2 mice (Myo15sh2/sh2). These mice have no functional myosin-XVa, which is critical for normal growth of mechanosensory stereocilia of hair cells. Although stereocilia of both inner and outer hair cells of Myo15sh2/sh2 mice lack myosin-XVa and are abnormally short, these cells have dramatically different hair bundle morphology. Myo15sh2/sh2 outer hair cells retain a staircase arrangement of the abnormally short stereocilia and prominent tip links. Myo15sh2/sh2 inner hair cells do not have obliquely oriented tip links, and their mechanosensitivity is mediated exclusively by "top-to-top" links between equally short stereocilia. In both inner and outer hair cells of Myo15sh2/sh2 mice, we found mechanotransduction responses with a normal "wild-type" amplitude and speed of activation. Surprisingly, only outer hair cells exhibit fast adaptation and sensitivity to extracellular Ca22+. In Myo15sh2/sh2 inner hair cells, fast adaptation is disrupted and the transduction current is insensitive to extracellular Ca22+.We conclude that the Ca22+ sensitivity of the mechanotransduction channels and the fast adaptation require a structural environment that is dependent on myosin-XVa and is disrupted in Myo15sh2/sh2 inner hair cells, but not in Myo15sh2/sh2 outer hair cells. [ABSTRACT FROM AUTHOR]
- Published
- 2009
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9. Auditory mechanotransduction in the absence of functional myosin-XVa.
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Stepanyan, Ruben, Belyantseva, Inna A., Griffith, Andrew J., Friedman, Thomas B., and Frolenkov, Gregory I.
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MYOSIN ,HAIR cells ,MECHANORECEPTORS ,AUDITORY pathways ,EAR diseases ,MORPHOGENESIS ,MEDICAL research - Abstract
In hair cells of all vertebrates, a mechanosensory bundle is formed by stereocilia with precisely graded heights. Unconventional myosin-XVa is critical for formation of this bundle because it transports whirlin and perhaps other molecular components responsible for programmed elongation of stereocilia to the stereocilia tips. A tip of a stereocilium is the site of stereocilia growth and one of the proposed sites of mechano-electrical transduction. In adult shaker 2 mice, a mutation that disables the motor function of myosin-XVa results in profound deafness and abnormally short stereocilia that lack stereocilia links, an indispensable component of mechanotransduction machinery. Therefore, it was assumed that myosin-XVa is required for proper formation of the mechanotransduction apparatus. Here we show that in young postnatal shaker 2 mice, abnormally short stereocilia bundles of auditory hair cells have numerous stereocilia links and ‘wild type’ mechano-electrical transduction. We compared the mechanotransduction current in auditory hair cells of young normal-hearing littermates, myosin-XVa-deficient shaker 2 mice, and whirler mice that have similarly short stereocilia but intact myosin-XVa at the stereocilia tips. This comparison revealed that the absence of functional myosin-XVa does not disrupt adaptation of the mechanotransduction current during sustained bundle deflection. Thus, the hair cell mechanotransduction complex forms and functions independently from myosin-XVa-based hair bundle morphogenesis. [ABSTRACT FROM AUTHOR]
- Published
- 2006
- Full Text
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10. Regulation of electromotility in the cochlear outer hair cell.
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Frolenkov, Gregory I.
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HAIR cells , *COCHLEA , *MAMMALS , *ACETYLCHOLINE , *CHLORIDE cells - Abstract
Mechanosensory outer hair cells play an essential role in the amplification of sound-induced vibrations within the mammalian cochlea due to their ability to contract or elongate following changes of the intracellular potential. This unique property of outer hair cells is known as electromotility. Selective efferent innervation of these cells within the organ of Corti suggests that regulation of outer hair cell electromotility may be the primary function of the efferent control in the cochlea. A number of studies demonstrate that outer hair cell electromotility is indeed modulated by the efferent neurotransmitter, acetylcholine. The effects of acetylcholine on outer hair cells include cell hyperpolarization and a decrease of the axial stiffness, both mediated by intracellular Ca2+. This article reviews these results and considers other potential mechanisms that may regulate electromotility, such as direct modification of the plasma membrane molecular motors, alteration of intracellular pressure, and modification of intracellular chloride concentration. [ABSTRACT FROM AUTHOR]
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- 2006
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11. Myosin-XVa is required for tip localization of whirlin and differential elongation of hair-cell stereocilia.
- Author
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Belyantseva, Inna A., Boger, Erich T., Naz, Sadaf, Frolenkov, Gregory I., Sellers, James R., Ahmed, Zubair M., Griffith, Andrew J., and Friedman, Thomas B.
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GREEN fluorescent protein ,HAIR cells ,CELLS ,RODENTS ,HEARING impaired ,MYOSIN - Abstract
Stereocilia are microvilli-derived mechanosensory organelles that are arranged in rows of graded heights on the apical surface of inner-ear hair cells. The 'staircase'-like architecture of stereocilia bundles is necessary to detect sound and head movement, and is achieved through differential elongation of the actin core of each stereocilium to a predetermined length. Abnormally short stereocilia bundles that have a diminished staircase are characteristic of the shaker 2 (Myo15a
sh2 ) and whirler (Whrnwi ) strains of deaf mice. We show that myosin-XVa is a motor protein that, in vivo, interacts with the third PDZ domain of whirlin through its carboxy-terminal PDZ-ligand. Myosin-XVa then delivers whirlin to the tips of stereocilia. Moreover, if green fluorescent protein (GFP)-Myo15a is transfected into hair cells of Myo15ash2 mice, the wild-type pattern of hair bundles is restored by recruitment of endogenous whirlin to the tips of stereocilia. The interaction of myosin-XVa and whirlin is therefore a key event in hair-bundle morphogenesis. [ABSTRACT FROM AUTHOR]- Published
- 2005
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12. Genetic insights into the morphogenesis of inner ear hair cells.
- Author
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Frolenkov, Gregory I., Belyantseva, Inna A., Friedman, Thomas B., and Griffith, Andrew J.
- Subjects
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MORPHOGENESIS , *INNER ear , *HAIR cells , *DEAFNESS , *MAMMALS , *GENETICS - Abstract
The mammalian inner ear is a sensory organ that has specialized hair cells that detect sound, as well as orientation and movement of the head. The 'hair' bundle on the apical surface of these cells is a mechanosensitive organelle that consists of precisely organized actin-filled projections known as stereocilia. Alterations in hair-bundle morphogenesis can result in hearing loss, balance defects or both. Positional cloning of genes that underlie hereditary hearing loss, coupled with the characterization of corresponding mouse models, is revealing how hair cells have adapted the molecular mechanisms of intracellular motility and intercellular adhesion for the morphogenesis of their apical surfaces. [ABSTRACT FROM AUTHOR]
- Published
- 2004
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13. Usher proteins in inner ear structure and function.
- Author
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Ahmed, Zubair M., Frolenkov, Gregory I., and Riazuddin, Saima
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INNER ear physiology , *USHER'S syndrome , *HEARING , *VISION , *HAIR cells , *ANIMAL models in research , *PATIENTS - Abstract
00135.2013.--Usher syndrome (USH) is a neurosensory disorder affecting both hearing and vision in humans. Linkage studies of families of USH patients, studies in animals, and characterization of purified proteins have provided insight into the molecular mechanisms of hearing. To date, 11 USH proteins have been identified, and evidence suggests that all of them are crucial for the function of the mechanosensory cells of the inner ear, the hair cells. Most USH proteins are localized to the stereocilia of the hair cells, where mechano-electrical transduction (MET) of sound-induced vibrations occurs. Therefore, elucidation of the functions of USH proteins in the stereocilia is a prerequisite to understanding the exact mechanisms of MET. [ABSTRACT FROM AUTHOR]
- Published
- 2013
- Full Text
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14. Nanoscale live-cell imaging using hopping probe ion conductance microscopy.
- Author
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Novak, Pavel, Chao Li, Shevchuk, Andrew I., Stepanyan, Ruben, Caldwell, Matthew, Hughes, Simon, Smart, Trevor G., Gorelik, Julia, Ostanin, Victor P., Lab, Max J., Moss, Guy W. J., Frolenkov, Gregory I., Klenerman, David, and Korchev, Yuri E.
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MICROSCOPY ,OPTICS ,NANOELECTROMECHANICAL systems ,MEDICAL microscopy ,HAIR cells - Abstract
We describe hopping mode scanning ion conductance microscopy that allows noncontact imaging of the complex three-dimensional surfaces of live cells with resolution better than 20 nm. We tested the effectiveness of this technique by imaging networks of cultured rat hippocampal neurons and mechanosensory stereocilia of mouse cochlear hair cells. The technique allowed examination of nanoscale phenomena on the surface of live cells under physiological conditions. [ABSTRACT FROM AUTHOR]
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- 2009
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15. RFX transcription factors are essential for hearing in mice.
- Author
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Elkon, Ran, Milon, Beatrice, Morrison, Laura, Shah, Manan, Vijayakumar, Sarath, Racherla, Manoj, Leitch, Carmen C., Silipino, Lorna, Hadi, Shadan, Weiss-Gayet, Michèle, Barras, Emmanuèle, Schmid, Christoph D., Ait-Lounis, Aouatef, Barnes, Ashley, Song, Yang, Eisenman, David J., Eliyahu, Efrat, Frolenkov, Gregory I., Strome, Scott E., and Durand, Bénédicte
- Subjects
DEAFNESS ,HAIR cells ,MECHANORECEPTORS ,SENSORY receptors ,KNOCKOUT mice - Abstract
Sensorineural hearing loss is a common and currently irreversible disorder, because mammalian hair cells (HCs) do not regenerate and current stem cell and gene delivery protocols result only in immature HC-like cells. Importantly, although the transcriptional regulators of embryonic HC development have been described, little is known about the postnatal regulators of maturating HCs. Here we apply a cell type-specific functional genomic analysis to the transcriptomes of auditory and vestibular sensory epithelia from early postnatal mice. We identify RFX transcription factors as essential and evolutionarily conserved regulators of the HC-specific transcriptomes, and detect Rfx1,2,3,5 and 7 in the developing HCs. To understand the role of RFX in hearing, we generate Rfx1/3 conditional knockout mice. We show that these mice are deaf secondary to rapid loss of initially well-formed outer HCs. These data identify an essential role for RFX in hearing and survival of the terminally differentiating outer HCs. [ABSTRACT FROM AUTHOR]
- Published
- 2015
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16. Mechanotransduction current is essential for stability of the transducing stereocilia in mammalian auditory hair cells.
- Author
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Lez-Ortega, A Catalina Vé, Freeman, Mary J., Indzhykulian, Artur A., Grossheim, Jonathan M., and Frolenkov, Gregory I.
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MECHANOTRANSDUCTION (Cytology) , *CALCIUM ions , *HAIR cells , *POTASSIUM antagonists , *INNER ear - Abstract
Mechanotransducer channels at the tips of sensory stereocilia of inner ear hair cells are gated by the tension of 'tip links' interconnecting stereocilia. To ensure maximal sensitivity, tip links are tensioned at rest, resulting in a continuous influx of Ca2+ into the cell. Here, we show that this constitutive Ca2+ influx, usually considered as potentially deleterious for hair cells, is in fact essential for stereocilia stability. In the auditory hair cells of young postnatal mice and rats, a reduction in mechanotransducer current, via pharmacological channel blockers or disruption of tip links, leads to stereocilia shape changes and shortening. These effects occur only in stereocilia that harbor mechanotransducer channels, recover upon blocker washout or tip link regeneration and can be replicated by manipulations of extracellular Ca2+ or intracellular Ca2+ buffering. Thus, our data provide the first experimental evidence for the dynamic control of stereocilia morphology by the mechanotransduction current. [ABSTRACT FROM AUTHOR]
- Published
- 2017
- Full Text
- View/download PDF
17. Expression of prestin, a membrane motor protein, in the mammalian auditory and vestibular periphery
- Author
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Adler, Henry J., Belyantseva, Inna A., Merritt Jr., Raymond C., Frolenkov, Gregory I., Dougherty, Gerard W., and Kachar, Bechara
- Subjects
- *
HAIR cells , *MAMMALS , *MECHANORECEPTORS , *PROTEINS - Abstract
Hair cells are specialized mechanoreceptors common to auditory and vestibular sensory organs of mammalian and non-mammalian species. Different hair cells are believed to share common features related to their mechanosensory function. It has been shown that hair cells possess various forms of motile properties that enhance their receptor function. Membrane-based electromotility is a form of hair cell motility observed in isolated outer hair cells (OHCs) of the cochlea. A novel membrane protein, prestin, recently cloned from gerbil and rat tissues, is presumably responsible for electromotility. We cloned prestin from mouse organ of Corti and confirmed strong homology of this protein among different rodent species. We explored whether or not prestin is present in hair cells of the vestibular system. Using reverse transcription-polymerase chain reaction, we demonstrated that prestin is expressed in mouse and rat auditory and vestibular organs, but not in chicken auditory periphery. In situ hybridization and immunolocalization studies confirmed the presence of prestin in OHCs as well as in vestibular hair cells (VHCs) of rodent saccule, utricle and crista ampullaris. However, in the VHCs, staining of varying intensity with anti-prestin antibodies was observed in the cytoplasm, but not in the lateral plasma membrane or in the stereociliary membrane. Whole-cell patch-clamp recordings showed that VHCs do not possess the voltage-dependent capacitance associated with membrane-based electromotility. We conclude that although prestin is expressed in VHCs, it is unlikely that it supports the form of somatic motility observed in OHCs. [Copyright &y& Elsevier]
- Published
- 2003
- Full Text
- View/download PDF
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