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逻辑后续，该工作显示了精神分裂症（SZ）和双相情感障碍（BD）受试者海马体（HIPP）固有神经回路的细微变化。其中一些异常表明，与细胞活力相关的通路中的细胞和分子变化可能导致bd和sz的细胞死亡，在较小程度上也可能导致sz的细胞死亡。我们最近使用基因表达谱（GEP）验证了这一假设，并对GenMapp生物途径/集群进行了新颖的事后分析。在bd中，与钙通道、单胺和肽g偶联蛋白受体（gpcr）以及包括凋亡、TFG-p和Wnt在内的多种信号通路相关的基因显著上调。在sz中，这些相同的通路受到影响，但方向相反，这表明至少其中一些变化可能与每种疾病的易感基因有关。为了进一步了解bd和SZs的HIPP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（SZs和bd）组成，称为McLean 66，另一个由SZs的一级亲属（SZ-F）和匹配的对照组成。在Specific Aim I中，将使用GEP、激光捕获显微解剖（LCM）和定量RT-PCR相结合的方法来确定这些变化是否存在于BDs和sz的HIPP中CA3/2和CA1区的特定亚区、层下和细胞亚型中。在以后的研究中，我们将在SZ-F队列中进行GEP，以确定sz中表达谱是否存在相似性。此外，我们的“部分”SZ啮齿动物模型的数据也将被用于评估BDs、SZ和pico处理大鼠中显示重叠变化的基因是否与HIPP的杏仁核激活有关。在Specific Aim II中，我们将通过双原位杂交（DISH）验证和扩展GEP结果，将与凋亡和其他GenMapp通路/簇相关的几个靶基因的mRNA与GAD65 mRNA共定位，以确定关键变化是否优先发生在gabaergy神经元中或非gabaergy神经元中。总的来说，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，这将提供更详细的了解基因表达的关键变化可能发生在HIPP电路的离散方面，以及这些变化是否可能与与精神障碍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.