Study of hair cell function and development in zebrafish

斑马鱼毛细胞功能和发育的研究

• 批准号: 6649034
• 负责人: Shuh-Yow Lin
• 金额: $ 4.81万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-12-02 至 2004-12-01
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6649034
• 关键词: auditory pathways; biological signal transduction; cell component structure /function; complementary DNA; developmental neurobiology; ear hair cell; electron microscopy; electrophysiology; gene expression; hearing; immunocytochemistry; in situ hybridization; mechanoreceptors; polymerase chain reaction; postdoctoral investigator; sensory mechanism; voltage /patch clamp; western blottings; zebrafish

Uncovering molecular mechanism of inner ear/lateral line hair cells functioning and development is the aim of this study. It will be among the first efforts to systematically screen through the zebrafish mutants generated by an insertional mutagenesis screen that is near completion in the Dr. Nancy Hopkins laboratory. This screen has identified over 500 genes which, when mutated, disrupts early development. Some of these genes have been identified as having a role in different part of nervous system development. These mutants will provide a fertile ground for identification of genes that participate the function and development of hair ceils. My first aim is to further characterize the pinball wizard (pwi) mutant, which have specific defects on mechanotransduction of hair cells. With the collaboration with Dr. David Corey's laboratory, electrophysiological analysis as well as morphological analysis at EM level will be used to gain further insight into the role of novel protein Pinball wizard (Pwi) in mechanotransduction. Second, I will screen through all of the mutants to identify those with a defect in styryl dye, FM 1-43, uptake. The Uptake of FM 1-43 has been shown to be pass directly through mechanotransduction channels at the tips of hair cell stereocilia. The mutants identified will either have impaired mechanotransduction, or have defects in hair cell development- They will be subject to further analysis, including a description of the mutant, the expression pattern of the gene during wild-type development.

这项研究的目的是发现内耳/侧线毛细胞的分子机制。这将是系统地筛选斑马鱼突变体，该突变体是由插入诱变屏幕所产生的，该突变体在南希·霍普金斯博士实验室即将完成。该屏幕已经确定了超过500个基因，这些基因在突变时会破坏早期发展。这些基因中的一些被确定为在神经系统发育的不同部分中起作用。这些突变体将为鉴定参与头发天花板功能和发展的基因提供肥沃的基础。我的第一个 目的是进一步表征弹球向导（PWI）突变体，该突变体具有特定的关于毛细胞机械转导的缺陷。通过与David Corey博士的实验室的合作，电生理分析以及EM水平的形态分析将用于进一步了解新型蛋白质弹球向导（PWI）在机械传输中的作用。其次，我将筛选所有突变体，以识别styryl染料缺陷的人，FM 1-43，摄取。 FM 1-43的摄取已被证明是直接通过毛细胞立体尾囊的机械传输通道传递的。确定的突变体将 机械转导障碍或毛细胞发育缺陷 - 它们将经过进一步的分析，包括对突变体的描述，野生型发育过程中基因的表达模式。