Calcium Signaling & Prefrontal Function in Schizophrenia

钙信号传导

• 批准号: 6892730
• 负责人: CLARE M BERGSON
• 金额: $ 16.45万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-12-12 至 2005-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6892730
• 关键词: animal tissue; behavioral genetics; calcium binding protein; calcium flux; dopamine; dopamine receptor; genetic susceptibility; genetically modified animals; human tissue; immunoelectron microscopy; immunoprecipitation; laboratory mouse; laboratory rat; mass spectrometry; matrix assisted laser desorption ionization; nerve /myelin protein; neural information processing; protein localization; protein protein interaction; proteomics; receptor expression; schizophrenia

Imbalances in cortical dopamine receptor (DR) mediated signaling are believed to underly both the cognitive and psychotic aspects of schizophrenia. Using yeast two-hybrid screens, we identified several DR-interacting proteins (DRIPs) that are prominently involved in intracellular calcium (Ca++) homeostasis (calcyon, NCS-1, TRPC-1, and CAPS). Disease-related increases in calcyon and NCS-1 protein levels were detected in dorsolateral prefrontal cortex (DLPFC) of patients with schizophrenia. We propose to extend our discovery based research on DR signaling complexes (DRSCs) guided by the hypothesis that abnormal dopamine-mediated Ca++ signaling is the basis of prefrontal dysfunction in schizophrenia. The possibility that this approach may also identify potential schizophrenia susceptibility genes (SSGs) is supported by a recent genetic association study which found evidence for an SSG at 10q26, the chromosomal position of the calcyon gene. We will utilize MALDI TOF-TOF mass spectrometry to identify the regional and receptor-specific composition of D1 and D2 DR-containing DRSCs immunoprecipitated from rat and monkey prefrontal cortex and striatum. It is likely that among the DRSC components, some proteins will interact directly with the receptor (DRIPs), while others (DRAPs or DR associated proteins) may interact with peripheral components of the complex. We will confirm interaction of DRSC proteins using a combination of biochemical and immunohistochemical methods. The effects of DRIP and DRAP interactions on DR function will be studied in transfected mammalian cells and native neural systems with a focus on Ca++ signaling and homeostasis. Expression of DRIPs and DRAPs will be analyzed in collections of schizophrenic brains. We will generate transgenic mice overexpressing NCS-1 or calcyon within the prefrontal cortex to learn whether these candidate schizophrenia-associated DRIPs are involved in the etiology of the disease. Transgenic mice will be subjected to a battery of behavioral, anatomical, and Physiological tests relevant to prefrontal deficits manifested in schizophrenia in collaboration with other subrojects. Identifying previously unknown proteins directly or indirectly associated with DRs should provide new insights into mechanisms of dopaminergic signaling in DLPFC, and clues as to the etiology of natomical and physiological defects manifested in schizophrenia.

皮质多巴胺受体（DR）介导的信号转导的不平衡被认为是精神分裂症的认知和精神病方面的基础。使用酵母双杂交筛选，我们确定了几个DR相互作用蛋白（DRIPs），主要参与细胞内钙（Ca++）稳态（calcyon，NCS-1，TRPC-1，和CAPS）。在精神分裂症患者的背外侧前额叶皮质（DLPFC）中检测到与疾病相关的钙蛋白和NCS-1蛋白水平增加。我们建议扩展我们的发现为基础的研究DR信号复合物（DRSC）的指导下的假设，即异常多巴胺介导的Ca++信号传导是前额叶功能障碍的精神分裂症的基础。最近的一项遗传关联研究支持了这种方法也可以识别潜在的精神分裂症易感基因（SSG）的可能性，该研究发现了在10 q26（calcyon基因的染色体位置）存在SSG的证据。我们将利用MALDI飞行时间-飞行时间质谱法，以确定区域和受体特异性组成的D1和D2 DR含有DRSCs免疫沉淀从大鼠和猴子的前额叶皮层和纹状体。很可能在DRSC组分中，一些蛋白质将直接与受体（DRIP）相互作用，而其他蛋白质（DRAP或DR相关蛋白）可能与复合物的外周组分相互作用。我们将使用生物化学和免疫组织化学方法的组合来确认DRSC蛋白的相互作用。DRIP和DRAP相互作用对DR功能的影响 将在转染的哺乳动物细胞和天然神经系统中进行研究，重点是Ca++信号传导和稳态。DRIP和DRAP的表达将在集合中进行分析 精神分裂症患者的大脑我们将产生转基因小鼠过度表达NCS-1或calcyon在前额叶皮层，以了解这些候选人精神分裂症相关的DRIP是否参与疾病的病因。将对转基因小鼠进行一系列行为、解剖学和生理学测试，这些测试与精神分裂症中表现出的前额叶缺陷有关，并与其他subrophylaxis合作。识别以前未知的蛋白质直接或间接与DR应DLPFC多巴胺能信号的机制提供新的见解，和线索的病因表现在精神分裂症的自然和生理缺陷。