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）介导的信号传导失衡被认为是精神分裂症认知和精神病方面的基础。使用酵母双杂交筛选，我们鉴定了几种主要参与细胞内钙 (Ca++) 稳态的 DR 相互作用蛋白 (DRIP)（calcyon、NCS-1、TRPC-1 和 CAPS）。在精神分裂症患者的背外侧前额皮质 (DLPFC) 中检测到与疾病相关的钙蛋白和 NCS-1 蛋白水平的增加。我们建议扩展我们对 DR 信号复合物 (DRSC) 的基于发现的研究，该研究以异常多巴胺介导的 Ca++ 信号传导是精神分裂症前额叶功能障碍的基础这一假设为指导。最近的一项遗传关联研究支持了这种方法还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 10q26（钙调蛋白基因的染色体位置）存在 SSG 的证据。我们将利用 MALDI TOF-TOF 质谱法来鉴定从大鼠和猴子前额皮质和纹状体免疫沉淀的含有 D1 和 D2 DR 的 DRSC 的区域和受体特异性组成。在 DRSC 成分中，一些蛋白质可能会直接与受体 (DRIP) 相互作用，而其他蛋白质（DRAP 或 DR 相关蛋白质）可能会与复合物的外周成分相互作用。我们将结合使用生化和免疫组织化学方法来确认 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。我们将产生在前额皮质内过度表达 NCS-1 或钙调蛋白的转基因小鼠，以了解这些候选精神分裂症相关 DRIP 是否与该疾病的病因有关。转基因小鼠将与其他子项目合作，接受一系列与精神分裂症中表现的前额叶缺陷相关的行为、解剖和生理学测试。鉴定以前未知的与 DR 直接或间接相关的蛋白质应该为 DLPFC 中多巴胺能信号传导机制提供新的见解，并为精神分裂症中表现的解剖和生理缺陷的病因学提供线索。