Calcium and Calmodulin Dependent Protein Kinase

钙和钙调蛋白依赖性蛋白激酶

• 批准号: 7156910
• 负责人: MARY B KENNEDY
• 金额: $ 31.97万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 1981
• 资助国家: 美国
• 起止时间: 1981-07-01 至 2008-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7156910
• 关键词: Actinin; Action Potentials; Acute; Alzheimer' s Disease; Appendix; Back; Birth; Brain; Calcium/calmodulin-dependent protein kinase; Chromosome Pairing; Complex; Cytoskeleton; Data; Defect; Development; Disease; Disruption; Docking; Enzyme Activation; Epilepsy; Genes; Glutamate Receptor; Glutamates; Goals; Heterozygote; Hippocampus (Brain); Knock-in Mouse; Knock-out; Knockout Mice; Kv4.2 channel; Learning; Link; Location; Maps; Mental Depression; Mitogen-Activated Protein Kinases; Molecular; Morphology; Mouse Protein; Mus; Mutant Strains Mice; N-Methyl-D-Aspartate Receptors; Neurologic; Neurons; Oral cavity; Pathology; Pathway interactions; Personal Satisfaction; Phenotype; Phosphorylation; Phosphotransferases; Physiological; Progress Reports; Prosencephalon; Protein Binding; Proteins; Psyche structure; Range; Recombinants; Recruitment Activity; Regulation; Regulatory Pathway; Research; Role; Schizophrenia; Signal Pathway; Signal Transduction; Site; Slice; Small Interfering RNA; Surface; Synapses; Synaptic Transmission; Synaptic plasticity; Tail; Testing; Vertebral column; Week; actin kinase; calmodulin-dependent protein kinase II; density; extracellular; falls; knock-down; mutant; postsynaptic; postsynaptic density protein; presynaptic density protein 95; protein activation; ras GTPase-Activating Proteins; receptor; synaptogenesis; tool

DESCRIPTION (provided by applicant): Derangements in synaptic transmission are part of the pathology of several neurological and mental diseases including epilepsy, schizophrenia, depression, and Alzheimer's disease. We are studying the molecular mechanisms underlying regulation of synaptic transmission. Here we propose to study regulatory pathways at glutamatergic synapses governed by Ca2+/calmodulin-dependent protein kinase II (CaMKII). CaMKII is concentrated in the postsynaptic density where it can be activated by Ca 2+ influx through NMDA receptors. The proposal focuses on two principal postsynaptic substrates of CaMKII, SynGAP, a Ras GTPase-activating protein that is concentrated in the postsynaptic density, and densin, a proposed docking site for CaMKII. In the First Aim, we will test the hypothesis that synGAP participates in regulation of the cytoskeleton at synapses in brain slices. We have previously shown that synGAP helps to regulate the spine cytoskeleton during synapse formation in cultured neurons, and that the location and activation of the kalirin/PAK kinase pathway is altered in hippocampal slices heterozygous for synGAP. We will use slices from wild type and synGAP deficient mutants to map the role of synGAP in this and related pathways following synaptic stimulation. In the Second Aim, we will use electrophysiological studies in hippocampal slices from wild type and conditional synGAP deficient mice to investigate whether the effects of synGAP deficiency on LTP are a result of developmental abnormalities or of acute loss of synGAP. We will also test the hypothesis that synGAP participates in regulation of the modulation of dendritic excitability by MAP kinase. In the Third Aim, we will examine recruitment of CaMKII to the PSD in neuronal cultures prepared from knock-in mice that are missing the carboxyl terminal tails of the NR2A and NR2B subunits of the NMDA receptor. We will also examine recruitment of CaMKII in these cultures after introducing recombinant forms of the intracellular tails of densin, or reducing the expression of densin by introduction of siRNA. In the Fourth Aim, we propose to determine the physiological importance of densin, and test the hypothesis that densin is a docking site for CaMKII in the PSD by constructing knockout and conditional knockout mutants of densin.

描述（由申请人提供）： 突触传递的紊乱是包括癫痫、精神分裂症、抑郁症和阿尔茨海默病在内的几种神经和精神疾病的病理学的一部分。我们正在研究突触传递调节的分子机制。在这里，我们建议研究的调节途径在海马神经元突触所管辖的钙/钙调蛋白依赖性蛋白激酶II（CaMKII）。CaMK Ⅱ主要集中在突触后致密区，可被NMDA受体介导的Ca 2+内流激活。该提案侧重于两个主要的突触后底物的CaMKII，SynGAP，Ras GTP酶激活蛋白，集中在突触后密度，和densin，一个建议的对接站点CaMKII。在第一个目标中，我们将测试synGAP参与调节脑切片中突触处的细胞骨架的假设。我们以前已经表明，synGAP有助于调节在培养的神经元突触形成过程中的棘细胞骨架，并且在synGAP杂合的海马切片中，kalirin/PAK激酶通路的位置和激活被改变。我们将使用野生型和synGAP缺陷突变体的切片来绘制synGAP在突触刺激后的这一通路和相关通路中的作用。在第二个目标中，我们将使用电生理研究野生型和条件synGAP缺陷小鼠海马切片，以调查是否synGAP缺陷对LTP的影响是由于发育异常或synGAP的急性损失。我们还将测试的假设，synGAP参与调节的调制树突兴奋的MAP激酶。在第三个目标中，我们将研究招聘的CaMK Ⅱ PSD的神经元培养物中制备的敲入小鼠是失踪的NMDA受体的NR 2A和NR 2B亚基的羧基末端的尾巴。我们还将研究在这些培养物中引入重组形式的致密蛋白的细胞内尾部后，或通过引入siRNA减少致密蛋白的表达后，CaMKII的募集。在第四个目标中，我们建议确定致密蛋白的生理重要性，并通过构建致密蛋白的敲除和条件敲除突变体来测试致密蛋白是PSD中CaMKII的对接位点的假设。