GENES CODING FOR INVERTEBRATE GAP JUNCTIONS

无脊椎动物间隙连接的基因编码

• 批准号: 6019448
• 负责人: ROBERT J WYMAN
• 金额: $ 27.54万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-08-01 至 2002-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6019448
• 关键词: Drosophilidae; Xenopus oocyte; electron microscopy; electrophysiology; gap junctions; gene expression; gene interaction; genetic polymorphism; in situ hybridization; injection /infusion; membrane channels; nucleic acid sequence; protein structure function; synapses; tissue /cell culture; transfection

We have defined a new gene family, the Passover family, whose members apparently code for the invertebrate gap junction channel. Thirty members sharing high sequence homology are known. Mutants are available for five members: mutations in three family members are viable and alter a few specific gap junctional connections; two genes are expressed widely and are mutable to lethality. The founding member of the family, Passover (Pas), has been expressed in Xenopus oocytes where it forms intercellular channels. This proposal seeks to start defining the properties of the junctions made by pas family genes. The genes will be expressed in specific cells in Drosophila by using the GALA-UAS system and in heterologous systems such as Xenopus oocytes and tissue culture cells by injection of RNA and by transfection. We will determine how different the channels made by different genes are and whether the different channel molecules form homotypic or heterotypic channels. In the animals, PAS is expressed in the neurons of the Giant Fiber system (GFS) which are connected by gap junctions. Pas mutations disconnect these cells. We will determine whether different transcripts rescue different synapses. Chimeric channel genes have been made and will be used to determine whether various aspects of function can be mapped to different molecular domains. A genetic screen will be undertaken to find molecules that interact with pas. Health significance: The pas family proteins have no known homologs in vertebrates. Drugs that target them specifically should be safe for human and vertebrate animals. This could lead to drugs that kill nematodes and other parasitic worms of plants and animals, These drugs should also be effective against insects, like the malaria mosquito, the tsetse fly, the deer tick, etc. for humans, as well as a host of ectoparasitic insects ( and probably mites) of veterinary importance. Of course, a prime target would be insects and other invertebrates that infest and east food crops. Identification of the molecules which form invertebrate gap junctions allows the methods of experimental crops. Identification of the molecules which form invertebrate gap junctions allows the methods of experimental manipulation available in flies and worms to be immediately useable for studies of gap junction communication. This should greatly facilitate studies of developmental mechanisms, channel function and neural systems.

我们定义了一个新的基因家族，逾越节家族，其成员 显然是无脊椎动物缝隙连接通道的编码。三十名成员 已知具有高序列同源性。突变体可供五人使用 成员：三个家庭成员的突变是可行的，并改变了一些 特定的间隙连接连接；两个基因广泛表达 是可变的致命性。家族创始成员逾越节 (Pas)，已在非洲爪蟾卵母细胞中表达，并形成细胞间质 渠道。该提案旨在开始定义 由 pas 家族基因形成的连接。 这些基因将通过使用以下方法在果蝇的特定细胞中表达： GALA-UAS 系统和异源系统，例如非洲爪蟾卵母细胞和 通过注射RNA和转染来组织培养细胞。我们将 确定不同基因产生的通道有多么不同， 不同通道分子形成同型还是异型 渠道。在动物中，PAS 在巨人神经元中表达 通过间隙连接连接的光纤系统（GFS）。 Pas突变 断开这些单元。我们将确定是否有不同的转录本 拯救不同的突触。嵌合通道基因已经制备并将 用于确定功能的各个方面是否可以映射到 不同的分子域。将进行基因筛查以发现 与 pa 相互作用的分子。 健康意义： pas 家族蛋白在 中没有已知的同源物 脊椎动物。专门针对它们的药物应该对人类是安全的 和脊椎动物。这可能会产生杀死线虫的药物 其他动植物寄生虫，这些药物也应 对昆虫有效，如疟蚊、采采蝇、 人类的鹿蜱等，以及许多体外寄生昆虫（ 可能还有螨虫）具有重要的兽医意义。当然是首要目标 可能是侵扰粮食作物的昆虫和其他无脊椎动物。 形成无脊椎动物间隙连接的分子的鉴定 允许试验作物的方法。分子的鉴定 形成无脊椎动物间隙连接允许实验方法 可在苍蝇和蠕虫中进行操作，可立即用于 间隙连接通讯的研究。这应该会极大地方便 发育机制、通道功能和神经系统的研究。