Genetic screens for analysis of Ca-dependent transmitter specification

用于分析 Ca 依赖性递质规范的遗传筛选

• 批准号: 7558888
• 负责人: NICHOLAS CANADAY SPITZER
• 金额: $ 28.04万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7558888
• 关键词: Acetylcholine; Action Potentials; Address; Area; Cells; Communities; Development; Embryo; Embryonic Development; Gene Expression; Genes; Genetic Screening; Glutamates; Glycine; Goals; Incidence; Insertional Mutagenesis; Length; Molecular; Mutation; Nature; Nervous System Physiology; Nervous system structure; Neurotransmitters; Process; Reagent; Regulation; Research; Role; Signal Pathway; To specify; Transgenic Organisms; Work; Xenopus; developmental disease; gain of function; gamma-Aminobutyric Acid; mutant; neuron development; public health relevance; research study

DESCRIPTION (provided by applicant): In previous work we demonstrated that Ca spikes generated by developmentally transient Ca-dependent action potentials regulate expression of neurotransmitters. We showed that this regulation was homeostatic, such that an increase in the Ca spike activity resulted in an increase in the incidence of cells expressing inhibitory neurotransmitters (glycine and GABA) and a concomitant decrease in the incidence of excitatory neurotransmitters (glutamate and acetylcholine). Conversely, a decrease in Ca spike activity caused an increase in the incidence of excitatory neurotransmitters and a decrease in inhibitory neurotransmitters. The proposed research has two specific aims targeted at generating mutant and transgenic lines in Xenopus tropicalis useful for identifying molecules involved in the Ca spike-dependent specification of neurotransmitters and for studying activity-dependent development in general. The first specific aim uses a gain-of-function screen and the second specific aim uses gene trap insertional mutagenesis. The third specific aim characterizes a subset of the genes identified in the first and second aims. The immediate goal of this research is to identify molecules involved in Ca spike- dependent neurotransmitter specification and to generate reagents for further research in this area. In addition, we aim to generate information and mutant lines that are of value to the Xenopus community. The long-term goal is to provide information about the molecular signaling pathways that govern processes of neuronal development, which will contribute to understanding developmental disorders of the nervous system. PUBLIC HEALTH RELEVANCE: Correct specification of neurotransmitter expression during embryonic development is essential for the function of the nervous system, and both gene expression and electrical activity contribute to this process. However the mechanisms by which nature (genes) and nurture (activity) interact to specify the appropriate expression of neurotransmitters are unclear. This research will identify activity-dependent genes regulating transmitter expression. It is expected that this work will contribute to understanding developmental disorders of the nervous system in which neurotransmitter expression is altered during embryonic and post-embryonic development.

描述（由申请人提供）： 在先前的工作中，我们证明了由发育短暂的CA依赖性作用电位产生的CA尖峰调节神经递质的表达。我们表明该调节是体内稳态的，因此CA尖峰活性的增加导致表达抑制性神经递质（甘氨酸和GABA）的细胞的发生率增加，并且兴奋性神经递质（谷氨酸和乙酰胆碱）的发生率伴随降低。相反，CA尖峰活性的降低导致兴奋性神经递质的发生率增加，并且抑制性神经递质的降低。拟议的研究具有两个特定目的，旨在在热带异植物中产生突变体和转基因线，这有助于识别涉及神经递质的CA尖峰依赖性规范和一般研究活动依赖性发展的分子。第一个特定的目的使用功能获得的屏幕，第二个特定目的使用基因陷阱插入诱变。第三个特定目的是在第一个和第二个目标中鉴定的基因的子集。这项研究的直接目的是确定涉及CA尖峰依赖性神经递质规范的分子，并生成试剂以在该领域进行进一步研究。此外，我们旨在生成对Xenopus社区有价值的信息和突变线。长期目标是提供有关控制神经元发展过程的分子信号通路的信息，这将有助于理解神经系统的发育障碍。公共卫生相关性：胚胎发育过程中神经递质表达的正确规范对于神经系统的功能至关重要，基因表达和电活动都有助于这一过程。但是，尚不清楚自然（基因）和培育（活性）相互作用以指定神经递质表达的机制。这项研究将确定调节发射器表达的活性依赖性基因。预计这项工作将有助于理解神经系统的发育障碍，在胚胎和胚胎后发育过程中，神经递质表达改变。