Si Fan, Brodie Hilary, Gillespie Peter G, Vazquez Ana E, Yamoah Ebenezer N
Center for Neuroscience, Department of Otolaryngology, University of California, Davis, Davis, California 95616, USA.
J Neurosci. 2003 Nov 26;23(34):10815-26. doi: 10.1523/JNEUROSCI.23-34-10815.2003.
Hair cells, the sensory receptors of auditory and vestibular systems, use a transducer apparatus that renders them remarkably sensitive to mechanical displacement as minute as 1 nm. To study the embryonic development of the transducer apparatus in hair cells of the chick auditory papilla, we examined hair cells that have been labeled with N-(3-triethylammoniumpropyl)-4-(4-(dibutylamino)styryl) pyridiniumdibromide, which has been shown to permeate the transducer channels. In addition, mechanotransduction currents were recorded directly using whole-cell patch-clamp techniques. The structure of the hair bundle was examined using scanning electron microscopy, and immunofluorescence labeling for myosin 1c, myosin 7a, and plasma membrane Ca2+ ATPase 2 was studied to determine the developmental expression of these proteins in embryonic chick papillas. We demonstrate that the transducer apparatus is assembled jointly at embryonic day 11 (E11) of the developing chick basilar papilla. The resting open probability of the transducer channels was high at E12 (approximately 0.5) and remained substantially elevated at E14-16; it then declined to the mature value of approximately 0.15 at E21. The displacement sensitivity of the transduction apparatus, the gating force, increased from E12 to E21. Although the expression of different components of the transducer apparatus and the transduction current peaked at approximately E14-16, marked refinement occurred beyond E16. For example, myosin 1c appeared diffusely localized in hair bundles from E12 to E16, but subsequently consolidated into punctate pattern. The fine temporal and precise spatial assembly of the transducer apparatus likely contributes toward the exquisite sensitivity of the transduction ensemble.
毛细胞是听觉和前庭系统的感觉感受器,它们使用一种换能器装置,使其对小至1纳米的机械位移具有极高的敏感性。为了研究鸡听觉乳头毛细胞中换能器装置的胚胎发育,我们检查了用N-(3-三乙铵丙基)-4-(4-(二丁基氨基)苯乙烯基)吡啶二溴化物标记的毛细胞,该物质已被证明可渗透换能器通道。此外,使用全细胞膜片钳技术直接记录机械转导电流。使用扫描电子显微镜检查毛束的结构,并研究肌球蛋白1c、肌球蛋白7a和质膜Ca2+ATP酶2的免疫荧光标记,以确定这些蛋白质在胚胎鸡乳头中的发育表达。我们证明,换能器装置在发育中的鸡基底乳头胚胎第11天(E11)共同组装。换能器通道的静息开放概率在E12时较高(约为0.5),并在E14 - 16时仍显著升高;然后在E21时降至约0.15的成熟值。转导装置的位移敏感性,即门控力,从E12到E21增加。尽管换能器装置不同组件的表达和转导电流在大约E14 - 16时达到峰值,但在E16之后发生了显著的优化。例如,肌球蛋白1c在E12到E16期间似乎分散地定位于毛束中,但随后合并成点状模式。换能器装置精细的时间和精确的空间组装可能有助于转导整体的极高敏感性。