MOLECULAR DELINEATION OF THE COCHLEAR HAIR CELLS

描述：(改编自《调查者摘要》)我们的长期目标是 了解耳蜗毛细胞作为有效频率的作用 外周听觉系统分析器。这项提议的目标是 确定电压门控离子通道超家族的成员 以确定耳蜗毛细胞的空间和时间模式 在发育和成体阶段的表达。这些研究将寻求 三叉神经节器官的解剖成熟与神经元神经支配的关系 CORTI与耳蜗毛细胞受体的功能成熟 潜在和突触传递。我们的研究将集中在人物塑造上 在分子水平上塑造受体电势的离子通道。 这些通道包括电压门控K+(Kv)和Na+(SCN)离子通道以及 内向整流器(KIR)。电压依赖性钙通道(CACN)和 参与耳蜗毛细胞的钙激活钾通道(Kca) 突触传递，也将被描述。初始基因表达 对这些离子通道的分析将使用退化基因家族和 用于从内耳中扩增cDNA的基因特异性寡核苷酸引物 毛细胞和外毛细胞的cDNA文库以及单细胞RT-PCR。 第一个目标是确定正常情况下毛细胞的离子通道 在分子水平上使用整体和切片的成年大鼠和小鼠 Corti器官的制备及对应答者的探查 蛋白质(免疫细胞化学)和信使核糖核酸(原位杂交)。一旦成虫 空间表达模式建立后，我们将研究时空关系 小鼠在产前和新生过程中的表达模式 确定这些离子通道的获取时间并关联这些变化 神经支配和解剖成熟。最后，我们将确定 缺失突变小鼠(NGN-1、NTRK2)离子通道表达的空间模式 (TrkB)、Ntf3(NT-3)、NTRK2+/Ntf3+/-和Ntf3+-PDGF启动子-Ntf3+)至 阐明神经支配与电生理的关系 成熟。受体相关分子元件的鉴定 耳蜗毛细胞的潜在和突触传递应导致 更好地了解内耳功能，有助于我们的理解 听觉病理学的结果。

DESCRIPTION: (Adapted from the Investigator's Abstract) Our long term goal is to understand how cochlear hair cells function as an effective frequency analyzer of the peripheral auditory system. The goals of this proposal are to identify members of the voltage-gated ion channel superfamily expressed in cochlear hair cells and to determine their spatial and temporal patterns of expression in developmental and adult stages. These studies will seek to correlate the anatomical maturation and neuronal innervation of the organ of Corti with the functional maturation of the cochlear hair cells' receptor potential and synaptic transmission. Our studies will focus on characterization of the ion channels that shape the receptor potential at the molecular level. These channels include the voltage-gated K+ (Kv) and Na+ (Scn) ion channels and the inward rectifiers (Kir). The voltage-dependent Ca2+ (CACN) channels and calcium-activated K+ channels (Kca), which are involved in cochlear hair cell synaptic transmissions, will also be characterized. Initial gene expression analysis of these ion channels will use degenerative gene-family and gene-specific oligonucleotide primers to amplify cDNAs from cochlear, inner hair cell, and outer hair cell cDNA libraries as well as single cell RT-PCR. The first aim will be to characterize the hair cells' ion channels in normal adult rats and mice at the molecular level using wholemount and section preparations of the organ of Corti and applying probes directed at candidate proteins (immunocytochemistry) and mRNA (in situ hybridization). Once the adult spatial expression patterns are established, we will study the spatio-temporal expression patterns in mice during a prenatal and neonatal time course to determine acquisition time of these ion channels and to correlate these changes with innervation and anatomical maturation. Lastly we will determine the spatial pattern of ion channel expression in null mutant mice (Ngn-1, Ntrk2 (trkB), Ntf3 (Nt-3), Ntrk2+/Ntf3+/-, and Ntf3+-PDGF promoter-Ntf3+) to elucidate the relationship between innervation and electrophysiological maturation. Identification of molecular elements involved in the receptor potential and synaptic transmission of cochlear hair cells should lead to a greater understanding of inner ear function and contribute to our comprehension of auditory pathologies.