The role of alpha2delta in neuronal VGCC modulation

alpha2delta 在神经元 VGCC 调节中的作用

• 批准号: 7902163
• 负责人: Cindy V Ly
• 金额: $ 2.44万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-01 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7902163
• 关键词: Absence Epilepsy; Affect; Alleles; Calcium; Calcium Channel; Cell membrane; Chemicals; Complex; Confocal Microscopy; Coupling; Defect; Drosophila genus; Drug or chemical Tissue Distribution; Excision; Exocytosis; Genes; Genetic; Goals; Homeostasis; Image; Immunohistochemistry; In Vitro; Induced Mutation; Lesion; Membrane; Membrane Protein Traffic; Molecular; Mus; Mutation; Nature; Neurons; Phenotype; Phosphorus; Play; Property; Regulation; Relative (related person); Role; Seizures; Severities; Staging; Synapses; System; Transducers; Vesicle; Work; in vivo; migration; mutant; neurotransmission; polyclonal antibody; stem; synaptogenesis; trafficking; voltage

DESCRIPTION (provided by applicant): The goal of this work is to better understand how auxiliary alpha2delta subunits modulate neuronal voltage-gated calcium channels (VGCCs) which affect neuronal migration (Komuro, et. al,1998), synaptogenesis (Bahls, et. al., 1998), and neurotransmission (Dunlap, 1995). Neuronal VGCCs consist of an alpha1 pore subunit and auxiliary subunits, alpha2delta, beta, and possibly gamma, which differentially regulate channel properties (Catterall, 2000). Studies in heterologous expression systems suggest that alpha2delta subunits associate with alpha1 and modulate channel function by altering the membrane trafficking and biophysical properties of VGCCs (Klugbauer, et. al, 2003). However, the in vivo function of alpha2delta in regulating pore-forming alpha1 subunits is less clear. Several straightjacket (stj) mutants with genetic lesions in the Drosophila alpha2delta subunit have been isolated. Initial electrophysiological and EM studies of stj mutants hint at defects in vesicle fusion. I propose to study the in vivo function of Drosophila alpha2delta, focusing on its role in regulating neuronal VGCCs and neurotransmission. Initially, I will generate null alleles of alpha2delta to aid in the genetic characterization of isolated mutations. I will then investigate VGCC trafficking in stj mutants using a fluorescent-tagged alpha1 pore subunit. In addition, I will determine whether alpha2delta modulates channel function using calcium imaging. Finally, I will explore whether interactions between alpha2delta and beta are necessary for proper VGCC function.

描述（由申请人提供）： 这项工作的目的是更好地了解辅助α2Delta亚基如何调节影响神经元迁移的神经元电压门控通道（VGCC）（Komuro等，等，1998），突触生成，突触发生（Bahls等）（Bahls等）（Bahls等）（Bahls等），和Neurotranss（Neurotrans），以及Dununlap，1995）。神经元VGCC由alpha1孔亚基和辅助亚基，alpha2delta，beta和可能的伽马组成，并可能调节通道性质（Catterall，2000）。在异源表达系统中的研究表明，通过改变VGCC的膜运输和生物物理特性，Alpha2delta亚基与Alpha1相关并调节通道功能（Klugbauer等人，等等，2003年）。但是，α2delta在调节孔形成α1亚基中的体内功能尚不清楚。已经分离出果蝇alpha2delta亚基中有几个具有遗传病变的直夹克（STJ）突变体。 STJ突变体的初始电生理学和EM研究暗示了囊泡融合中的缺陷。我建议研究果蝇α2delta的体内功能，重点是其在调节神经元VGCC和神经传递中的作用。最初，我将产生α2Delta的无效等位基因，以帮助分离突变的遗传表征。然后，我将使用荧光标记的alpha1孔亚基研究VGCC贩运STJ突变体。此外，我将确定α2Delta是否使用钙成像调节通道函数。最后，我将探讨alpha2delta和beta之间的相互作用是否对于适当的VGCC功能是必需的。