NEUROGENETICS OF SODIUM CHANNEL GENES IN DROSOPHILA

果蝇钠通道基因的神经遗传学

基本信息

批准号：
2684920
负责人：
BARRY S GANETZKY
金额：
$ 17.23万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
1989
资助国家：
美国
起止时间：
1989-12-01 至 1999-03-31
项目状态：
已结题

项目摘要

DESCRIPTION: Na channels perform the central role in the generation and propagation of action potentials, which are the primary form of electrical signalling in the nervous system. The long term objective of the proposed work is to elucidate the molecular mechanisms that underlie neural signalling. Here, the investigator focuses on analysis of the structure, function, and regulation of two Na channel structural genes in Drosophila, called para and Dsc. These Na channel genes comprise a small family whose members appear to be differentially utilized and could subserve physiologically distinct functions. The goals are to understand the respective functions of these two Na channel genes in the Drosophila nervous system, the basis of their differential expression, and the phenotypic consequences of mutations. The investigator will approach these goals using a combination of genetic, molecular, and electrophysiological techniques. To characterize Dsc, cDNAs representing the entire open reading frame will be isolated and sequenced; null mutations will be generated by imprecise excision of P element insertions within the gene; and behavioral and electrophysiological phenotypes of these mutants alone and in various double mutant combinations will be characterized. To characterize further the spatial and developmental expression of para and Dsc and to elucidate some of the regulatory mechanisms governing their expression, the investigator will: raise para- and Dsc-specific antisera for immunolocalization of the corresponding channel polypeptides; examine the cellular distribution of the large array of para splice forms using exon- specific probes for in situ hybridization, splice-dependent reporter constructs, or single-cell PCR analysis; and identify the upstream transcriptional regulatory sequences of para for use in germline transformation experiments. To explore structure-function relationships of para and Dsc, the investigator will determine the exact lesion in a large collection of in vivo-generated para mutations using a sensitive SSCP technique in combination with sequence analysis. Electrophysiological recording of Na currents in cultured Drosophila neurons and in heterologous expression systems will be used to compare the properties of channels encoded by wild-type and mutant alleles of para and Dsc as well as by the different splice variants of para. Finally, the investigator proposes to use a PCR-based strategy to search for additional Na channel genes in Drosophila that will be targeted for mutagenesis and phenotypic analysis. Because para and Dsc are the only neuronal Na channel genes in any organism that have been mutated in situ, these genes provide unique opportunities to obtain novel insights into the molecular mechanisms of Na channel expression, function, and regulation, in vivo. A number of human hereditary neuromuscular diseases are known to be associated with perturbation in the structure or function of ion channels, including Na channels. Therefore, the results obtained from these studies could have direct significance for the understanding and possible treatment of these disorders.

描述：Na 通道在生成和动作电位的传播，这是动作电位的主要形式神经系统中的电信号。长期目标拟议工作的目的是阐明分子机制是神经信号传导的基础。在这里，研究者重点分析两个Na通道结构的结构、功能和调控果蝇中的基因，称为 para 和 Dsc。这些Na通道基因由一个小家庭组成，其成员似乎有所不同利用并可以促进生理上不同的功能。这目标是了解这两个Na通道各自的功能果蝇神经系统中的基因，其差异的基础表达以及突变的表型后果。这研究人员将结合遗传、分子和电生理学技术。为了表征 Dsc，代表整个开放阅读框的 cDNA 将被分离并测序； P的不精确切除将产生无效突变基因内的元件插入；以及行为和这些突变体单独和不同的电生理表型双突变体组合将被表征。表征进一步增强 para 和 Dsc 的空间和发展表达，并阐明一些控制其表达的调节机制，研究者将：产生 para 和 Dsc 特异性抗血清相应通道多肽的免疫定位；检查使用外显子对大量对剪接形式的细胞分布用于原位杂交、剪接依赖性报告基因的特异性探针构建体或单细胞 PCR 分析；并确定上游用于种系的para转录调控序列转化实验。探索结构与功能的关系 para 和 Dsc 中，研究者将确定 a 中的确切病变使用敏感的方法收集大量体内产生的副突变 SSCP 技术与序列分析相结合。培养果蝇中Na电流的电生理记录神经元和异源表达系统将用于比较野生型和突变型等位基因编码的通道的特性 para 和 Dsc 以及 para 的不同剪接变体。最后，研究者建议使用基于PCR的策略来搜索果蝇中额外的 Na 通道基因将被靶向诱变和表型分析。因为 para 和 Dsc 是唯一的任何生物体中的神经元Na通道基因已发生突变在原位，这些基因提供了获得新见解的独特机会研究 Na 通道表达、功能和的分子机制调节，体内。多种人类遗传性神经肌肉疾病已知与结构中的扰动有关或离子通道的功能，包括Na通道。因此，结果从这些研究中获得的结果可能对对这些疾病的了解和可能的治疗。

项目成果

期刊论文数量（13）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Molecular characterization of neurally expressing genes in the para sodium channel gene cluster of drosophila.

果蝇副钠通道基因簇中神经表达基因的分子特征。

DOI：
10.1093/genetics/142.3.879
发表时间：
1996
期刊：
Genetics
影响因子：
3.3
作者：
Hong,CS;Ganetzky,B
通讯作者：
Ganetzky,B

A glial-neuronal signaling pathway revealed by mutations in a neurexin-related protein.

神经毒素相关蛋白突变揭示的神经胶质神经元信号通路。

DOI：
10.1126/science.283.5406.1343
发表时间：
1999
期刊：
Science (New York, N.Y.)
影响因子：
0
作者：
Yuan,LL;Ganetzky,B
通讯作者：
Ganetzky,B

Genetic analysis of ion channels in Drosophila.

果蝇离子通道的遗传分析。

DOI：
发表时间：
1991
期刊：
Epilepsy research. Supplement
影响因子：
0
作者：
Ganetzky,B
通讯作者：
Ganetzky,B

Functional expression of Drosophila para sodium channels. Modulation by the membrane protein TipE and toxin pharmacology.

DOI：
10.1085/jgp.110.2.119
发表时间：
1997-08
期刊：
JOURNAL OF GENERAL PHYSIOLOGY
影响因子：
3.8
作者：
Warmke, J W;Reenan, R A;Wang, P;Qian, S;Arena, J P;Wang, J;Wunderler, D;Liu, K;Kaczorowski, G J;Van der Ploeg, L H;Ganetzky, B;Cohen, C J
通讯作者：
Cohen, C J

Conserved alternative splicing patterns and splicing signals in the Drosophila sodium channel gene para.

果蝇钠通道基因中保守的选择性剪接模式和剪接信号。