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，显示了精神分裂症（SZ）和双相情感障碍（BD）受试者海马（HIPP）内在神经回路的细微变化。这些异常中的一些表明，与细胞活力相关的途径中的细胞和分子变化可能导致BD中的细胞死亡，并且在较小程度上导致SZ中的细胞死亡。我们最近使用基因表达谱（GEP）以及GenMapp生物通路/簇的新事后分析来验证这一假设。在BD中观察到与钙通道、单胺和肽G偶联蛋白受体（GPCR）以及包括凋亡、TFG-p和Wnt在内的多种信号传导途径相关的基因的显著上调。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。为了更多地了解BD和SZ的HIPP中的细胞调节机制，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BD组成，称为姆克林66，另一个由SZ的一级亲属（SZ-F）和匹配的对照组成。在特定目标I中，GEP、激光捕获显微切割（LCM）和定量RT-PCR的组合将用于确定这些变化是否存在于BD和SZ的HIPP中CA 3/2与CA 1扇区的特定亚区、亚板层和细胞亚型中。在以后的研究中，我们将在SZ-F队列中进行GEP，以确定SZ中的表达谱是否存在相似性。此外，从我们的“部分”啮齿动物模型的SZ，其中印防己毒素是急性和慢性注入杏仁核的数据也将被用来评估是否在BD，SZ和PICRO处理的大鼠的基因显示重叠的变化可能与杏仁核激活的HIPP。在特定目标II中，我们将通过使用双重原位杂交（DISH）将来自与细胞凋亡相关的几个靶基因和其他GenMapp通路/簇的mRNA与GAD 65 mRNA共定位，以确定关键变化是否优先发生在GABA能或非GABA能神经元中，从而验证和扩展GEP结果。总的来说，拟议的研究正在寻求复制，验证和扩展我们的GEP研究的关键方式，将提供一个更详细的了解基因表达的关键变化可能发生在离散方面的HIPP电路，以及这种变化是否可能与遗传或环境因素相关的精神疾病。

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.