Molecular & Biochemical Core

额叶皮质、纹状体、丘脑和杏仁核内神经元回路的异常是精神分裂症病理生理状态的主要组成部分。由于不可能研究特定的神经元群体，抗精神病药物的分子机制的鉴定受到了极大的影响。Conte中心建议通过使用一种新技术来解决这个问题，该技术可以精确地分析复杂混合和解剖的个别类型的神经元 难以辨别。这种多学科方法涉及许多可以在诸如核心这样的中央结构中有效执行的重复性任务。在过去的20年里，格林加德实验室在运营这样一个设施方面积累了相当多的经验。核心的一个重要方面将是支持确定的新候选基因的特征。这些职责将包括关键试剂库存的生产和维护、新试剂的开发以及例行任务的执行。具体目标包括： I.多克隆/单克隆抗体的产生、制备、纯化和鉴定，包括磷酸化状态特异性抗体。核心将通过产生各种多克隆和单克隆抗体来支持整个计划。此外，还将根据需要生产、测试和提纯针对关键信号分子的磷酸特异性抗体。 项目1-5所要求的病毒试剂的设计和生产。Core将设计和生产必要的AAV病毒试剂，以推动在特定细胞群中表达Cre重组酶的小鼠品系中siRNAs或蛋白质的特定表达。 三、例行任务的执行情况。这将包括各种激酶、磷酸酶和细胞内信号分子的制备、纯化和表征。Core将在需要的情况下进行酵母双杂交研究。核心人员还在各种其他常规任务方面拥有丰富的经验，如细胞培养准备、免疫荧光技术、siRNA设计和测试。

Abnormalities in neuronal circuits within frontal cortex, striatum, thalamus and amygdala represent a major component of the pathophysiologic state of schizophrenia. The identification of the molecular mechanisms of antipsychotic drugs has been greatly impaired by the impossibility of studying specific neuronal populations. This Conte Center proposes to address this issue by using a novel technology that precisely allows the analysis of individual types of neurons that are intricately mixed and anatomically indistinguishable. This multidisciplinary approach involves a number of repetitive tasks that can be efficiently conducted in a centralized structure such as a Core. Over the past 20 years, considerable experience has been acquired in the Greengard laboratory with the operation of such a facility. An important aspect of the Core will be to support the characterization of novel candidate genes identified. Such responsibilities will include the production and maintenance of key reagents stocks, the development of new reagents, and the performance of routine tasks. Specific Aims include: I. The generation, production, purification and characterization of polyclonal/monoclonal antibodies, including phosphorylation state-specific antibodies. The Core will support the entire program by generating various polyclonal and monoclonal antibodies. In addition, phospho-specific antibodies against key signaling molecules will be produced, tested, and purified as needed. II. The engineering and production of viral reagents required by Projects 1-5. The Core will design and generate the necessary AAV viral reagents in order to drive specific expression of siRNAs or protein overexpression in mouse lines that express the Cre recombinase in specific cell populations. III. The performance of routine tasks. This will include the preparation, purification and characterization of various kinases, phosphatases, and intracellular signaling molecules. The Core will perform, where needed, yeast two-hybrid studies. The Core personnel has also extensive experience in various other routine tasks such as cell culture preparation, immunofluorescence techniques, siRNA design and testing.