Gene Expression in Hippocampal Circuits of Bipolars and Schizophrenics

描述(由申请人提供)：本提案中描述的研究是我们早期工作的合乎逻辑的后续，显示了精神分裂症(SZ)和双相情感障碍(BD)受试者海马体(HIPP)内在神经回路的细微变化。其中一些异常提示，与细胞活性相关的细胞和分子通路的变化可能导致BDS和SZ的细胞死亡，程度较轻。我们最近使用基因表达谱(GEP)，结合对GenMapp生物路径/簇的新的后期分析，验证了这一假设。在BDS中，与钙通道、单胺和多肽G偶联蛋白受体(GPCRs)相关的基因以及包括细胞凋亡、TFG-p和Wnt在内的多种信号通路都有明显的上调。在SZ中，这些相同的通路受到影响，但方向相反，这表明这些变化中至少有一些可能与每种疾病的易感基因有关。为了更多地了解BDS和SZ的HIPP的细胞调控机制，我们建议使用两个队列，一个由一组特征良好的正常对照、SZ和BDS组成，称为McLean 66，另一个由SZ的一级亲属(SZ-F)和匹配的对照组组成。在具体目标I中，将结合GEP、激光捕获显微解剖(LCM)和定量RT-PCR来确定这些变化是否存在于BDS和SZ的HIPP的CA3/2和CA1扇区的特定亚区、亚胺和细胞亚型。在以后的研究中，我们将在SZ-F队列中进行GEP，以确定SZ的表达谱是否有相似性。此外，我们的SZ“部分”啮齿动物模型中的数据也将被用来评估BDS、SZ和PICRO治疗的大鼠中显示重叠变化的基因是否可能与杏仁核Hipp的激活有关。在特定的目标II中，我们将通过使用双重原位杂交(DISH)来验证和扩展GEP结果，以确定关键变化是否优先发生在GABA能或非GABA能神经元中，以确定与细胞凋亡相关的几个靶基因以及其他GenMapp通路/簇与GAD65 mRNA的共定位。总体而言，拟议的研究正在寻求以关键的方式复制、验证和扩展我们的GEP研究，以提供更详细的了解，以了解在HIPP回路的不同方面基因表达的关键变化可能发生在哪里，以及这些变化是否可能与与精神病相关的遗传或环境因素有关。

DESCRIPTION (provided by applicant): The studies described in this proposal are a logical follow-up to our earlier work showing subtle alterations of intrinsic neural circuitry in the hippocampus (HIPP) of schizophrenic (SZ) and bipolar disorder (BD) subjects. Some of these abnormalities have suggested that cellular and molecular changes in pathways associated with cell viability may lead to cell death in BDs and, to a lesser degree, SZs. We recently tested this hypothesis using gene expression profiling (GEP), together with a novel post hoc analysis of GenMapp biopathways/clusters. A pronounced upregulation of genes associated with calcium channels, monoamine and peptide G-coupled protein receptors (GPCRs) and multiple signaling pathways including apoptosis, TFG-p and Wnt has been observed in BDs. In SZs, these same pathways are affected, but in the opposite direction, suggesting that at least some of these changes may be related to susceptibility genes for each of the disorders. To learn more about cellular regulatory mechanisms in the HIPP of BDs and SZs, we are proposing to use two cohorts, one composed of a well-characterized set of normal controls, SZs and BDs called the McLean 66 and another consisting of first degree relatives of SZs (SZ-F) and matched controls. In Specific Aim I, a combination of GEP, laser capture microdissection (LCM) and quantitative RT-PCR will be used to determine whether these changes are present in particular subregions, sublaminae and cellular subtypes in sectors CA3/2 versus CA1 in HIPP of BDs and SZs. In later studies, we will perform GEP in the SZ-F cohort to determine whether there are similarities in the expression profiles in SZs. In addition, data from our 'partial' rodent model of SZ in which picrotoxin is acutely and chronically infused in the amygdala will also be used to evaluate whether genes showing overlapping changes in BDs, SZs and PICRO-treated rats may be related to amygdalar activation of the HIPP. In Specific Aim II, we will validate and extend the GEP results by using double in situ hybridization (DISH) to co-localize mRNA from several target genes associated with apoptosis and other GenMapp pathways/clusters with GAD65 mRNA to determine whether key changes are preferentially occurring in GABAergic or non-GABAergic neurons. Overall, the proposed studies are seeking to replicate, validate and extend our GEP studies in critical ways that will provide a more detailed understanding of where pivotal changes in gene expression may be occurring within discrete aspects of HIPP circuitry and whether such changes may be related to genetic or environmental factors associated with the psychotic disorders.