GENETICS OF NEMATODE PHARYNGEAL MUSCLE EXCITABILITY

这项研究的广泛和长期目标是开发一种 无脊椎动物模型系统，允许研究心脏的兴奋性， 就像肌肉一样，通过一系列的药理学，电生理学， 分子和遗传技术，重点是分子遗传学。 该模型将用于识别在兴奋性中重要的分子， 这可能有助于理解疾病（例如，心律失常）， 心脏兴奋性的缺陷，以及理解和 开发治疗这些疾病的药物。 该模型 秀丽隐杆线虫的咽肌。 a）、 影响咽肌兴奋性的基因的鉴定。 将分离出咽肌异常的突变体 B）eat-5的遗传分析，其影响咽部的耦联。 c）对eat-6的遗传分析，可能是正常肌肉所必需的。 咽肌复极 将使用经典的遗传学方法 为了确定咽肌的行为如何受到完全丧失 eat-5或eat-6的功能，基因何时起作用，以及它们如何相互作用; d） eat-5的分子克隆和表征; EAT-6的表征。 RFLP和缺失作图将用于 将线虫基因组的遗传图谱和物理图谱关联起来， EAT-5和EAT-6附近。 这些基因将通过它们的能力 来拯救变异蠕虫的表型 cDNA将被分离， f）功能的药理学表征 孤立的咽喉 咽部将被解剖出蠕虫和他们的 体外研究功能。

The broad, long-term objective of this study is the development of an invertebrate model system allowing the study of excitability of a heart- like muscle by the full array of pharmacological, electrophysiological, molecular, and genetic techniques, with the emphasis on molecular genetics. This model will be used to identify molecules important in excitability, which may help in understanding diseases (e.g., arrhythmias) that can result from defects in cardiac excitability, and in understanding and developing pharmacological agents to treat these diseases. The model is the pharyngeal muscle of the nematode Caenorhabditis elegans. a) Identification of genes that affect pharyngeal muscle excitability. Mutants will be isolated that show abnormalities in pharyngeal muscle motions; b) Genetic analysis of eat-5, which affects coupling of pharyngeal muscles; c) Genetic analysis of eat-6, probably necessary for normal pharyngeal muscle repolarization. Classical genetic methods will be used to determine how pharyngeal muscle behavior is affected by complete loss of eat-5 or eat-6 function, when the genes act, and how they interact; d) Molecular cloning and characterization of eat-5; e) Molecular cloning and characterization of eat-6. RFLP and deletion mapping will be used to correlate the genetic and physical maps of the C elegans genome in the vicinity of eat-5 and eat-6. The genes will be identified by their ability to rescue the phenotype of mutant worms. cDNAs will be isolated and sequenced, and f) Pharmacological characterization of the function of isolated pharynxes. Pharynxes will be dissected out of worms and their function studied in vitro.