PROMOTER CHARACTERIZATION OF THE CART GENE

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 这是我们实验室的主要研究项目，其中CART基因表达的调控一直是重点。我们以前发现CREB在N-ACC中的过度表达增加了CART多肽的表达，我们发现CREB在体内是CART表达的调节因子，这似乎是可卡因作用的机制之一。CART肽起到体内平衡调节剂的作用，并倾向于对抗可卡因在伏隔核的作用。在过去的一段时间里，我们使用芯片分析表明，CREB和P-CREB与CART启动子中包含Cre位点的区域结合，表明CREB对CART基因有直接影响。 支持的其他工作是鉴定在体内注射后降低CART多肽水平的siRNAs。已经发现，在初步实验中，有几个siRNA可以降低CART多肽水平。这一点很重要，因为到目前为止，还没有办法急剧和突然地降低大脑区域的CART多肽水平。如果CART的多肽水平能够显著降低，就可以进行有用的行为研究。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. This is the main research project in our laboratory where regulation of CART gene expression has been a focus. We previously showed that over expression of CREB in the N Acc increased expression of CART peptide and we found that CREB is an in vivo regulator of CART expression which appears to be a mechanism of the action of cocaine. CART peptide acts as a homeostatic regulator and tends to oppose the action of cocaine in the nucleus accumbens. In the past period, we have shown using ChIP assays that CREB and P-CREB bind to the region of the CART promoter that contains the CRE site, suggesting a direct effect of CREB on the CART gene. Additional work supported has been the identification of siRNAs that reduce CART peptide levels after in vivo injection. Several siRNAs have been found that, in preliminary experiments, reduce CART peptide levels. This is important because until now there has been no way to acutely and abruptly reduce CART peptide levels in brain regions. Useful behavioral studies can be carried out if CART peptide levels can be acutely reduced.