Somatic Ca2+ Signaling in CNS Neurons

CNS 神经元中的体细胞 Ca2 信号传导

• 批准号: 6987888
• 负责人: DONNA L GRUOL
• 金额: $ 33.91万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-01-01 至 2007-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6987888
• 关键词: biological signal transduction; calcium flux; calcium ion; central nervous system; cerebellar Purkinje cell; developmental genetics; gene expression; microarray technology; neurogenesis; neurogenetics

DESCRIPTION (provided by applicant): Neuronal activity produces dynamic changes in cellular calcium that play a central role in the regulation of a number of cellular functions including gene expression. Recently we demonstrated that early developing cerebellar Purkinje neurons express a membrane-to-nucleus somatic calcium signaling pathway that is driven by endogenously generated electrical activity at a developmentally significant stage, just prior to dendritic expression. Our preliminary results show that activation of this signaling pathway produces cytosolic and nuclear calcium signals and that these signals result in activation of transcription factors and changes in the level of downstream cellular proteins. We hypothesize that the endogenously generated activity that drives this pathway in the immature Purkinje neurons is a developmentally relevant signal and that one role for the activity is to induce changes in gene expression necessary for initiation of the next developmental stage, expression of functional dendrites. As a first step toward testing this hypothesis, we will apply gene array microanalysis to acutely isolated Purkinje neurons subjected to various stimulation paradigms at the relevant developmental stage. This approach will enable us to simultaneously analyze the effects of neuronal activation on a large number of neuronal transcripts known to contribute to neuronal function and development. By using this information in combination with analysis of protein expression and functional analysis using electrophysiological and calcium imaging techniques we will be able to gain an understanding of the impact of calcium-regulated gene expression at a specific developmental stage. We will also identify the molecular components and functional properties of the membrane-to-nucleus somatic calcium signaling pathway and determine the conditions that are essential for gene expression of proteins identified by the gene array microanalysis. Results from these studies will be directly relevant to the developmental program of Purkinje neurons, which has been well characterized on a morphological level and involves distinct and identifiable developmental stages. However, the general principles identified by the studies will also contribute significantly to our understanding of the role of activity-dependent gene expression during development in other CNS neuronal types.

描述（由申请人提供）： 神经元活性会在细胞钙中产生动态变化，在调节许多细胞功能（包括基因表达）中起着核心作用。最近，我们证明了早期发育的小脑Purkinje神经元表达膜至核核钙信号传导途径，该通路是由内源性产生的电活动在发育意义的阶段驱动的，就在树突状表达之前。我们的初步结果表明，该信号通路的激活会产生胞质和核钙信号，并且这些信号导致转录因子激活以及下游细胞蛋白水平的变化。我们假设，在未成熟的浦肯野神经元中驱动这种途径的内源性生成的活性是一个发育相关的信号，该活动的一种作用是诱导启动下一个发育阶段，功能性树突表达所必需的基因表达变化。作为检验这一假设的第一步，我们将在相关发育阶段将基因阵列微分析应用于急性分离的Purkinje神经元。这种方法将使我们能够同时分析神经元激活对已知有助于神经元功能和发育的大量神经元转录本的影响。通过将这些信息与使用电生理和钙成像技术的蛋白质表达和功能分析的分析结合使用，我们将能够了解在特定发育阶段钙调节基因表达的影响。我们还将确定膜到核体细胞钙信号通路的分子成分和功能特性，并确定基因阵列微分析鉴定的蛋白基因表达所必需的条件。这些研究的结果将与Purkinje神经元的发展计划直接相关，后者在形态学层面上得到了很好的特征，并且涉及独特且可识别的发育阶段。但是，研究确定的一般原则还将显着有助于我们对其他中枢神经系统神经元类型发育中活性依赖性基因表达的作用的理解。