GENETICS OF NEMATODE PHARYNGEAL MUSCLE EXCITABILITY

这项研究的广泛和长期目标是发展一种 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，由一整套的药理、电生理、 分子和遗传技术，重点是分子遗传学。 这个模型将被用来识别对兴奋性很重要的分子， 这可能有助于了解疾病(例如心律失常) 由于心脏兴奋性的缺陷，以及理解和 开发治疗这些疾病的药理药剂。该模型是 线虫线虫的咽部肌肉。a) 识别影响咽肌兴奋性的基因。 将分离出显示咽部肌肉异常的突变体 运动；b)影响咽部连接的EAT-5基因分析 肌肉；c)Eat-6的基因分析，可能是正常所必需的 咽肌复极。将使用经典的遗传方法 为了确定咽肌行为如何受到完全丧失的影响 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；e)分子克隆和鉴定 EAT-6的特性。将使用RFLP和删除映射来 将线虫基因组的遗传图谱和物理图谱联系起来 在Eat-5和Eat-6附近。这些基因将通过它们的能力来识别 以挽救突变蠕虫的表型。CDNA将被分离并 测序，和f)功能的药理学特征 孤立的咽部。咽部将被解剖出蠕虫和它们的 体外功能研究。

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.