基于microRNAs与Glu-NMDAR神经递质系统在甲基苯丙胺依赖中的分子机制研究

我们前期鉴定的对神经系统有影响的CACNA1C基因被证实是miR-137靶基因，且CACNA1C编码的Ca(v)1.2被发现在药物依赖中参与Glu-NMDAR神经递质系统和可能作用于ΔFosB。本项目系统研究基于miRNAs的MA依赖分子机制。首先通过Solexa测序和芯片建立MA敏化小鼠和正常小鼠全基因组miRNAs和mRNAs差异表达谱。其次，运用生物信息学和Q-PCR等方法，在另一独立样本中明确关键调控miRNAs及其可能的靶基因。再次，对确定的关键调控miRNAs靶基因进行生物学验证，并利用报告基因和ChIP等方法明确关键调控miRNAs靶基因及上游调控机制。最后，利用MA敏化小鼠模型通过RNAi等技术在体上/下调靶基因的表达，阐明miRNAs及其靶基因对Glu-NMDAR神经递质系统及ΔFosB作用的分子机制。本研究有助于拓展对MA依赖分子机制的认识并为其研究提供新线索和新思路。

CACNA1C which effects on the nervous system was proven to be the target gene of miR-137 in previous experiments and its encoding product, Ca(v)1.2, might involve in Glu-NMDAR neurotransmitter system and act on ΔfosB. The aim of this research is to explore the molecular mechanism of methamphetamine addiction based on miRNAs. Firstly, we will establish the differential expression profile of the whole-genome miRNAs and mRNAs of sensitized and normal mice by Solexa sequencing and chips. Secondly, we will determine the critical regulated miRNAs and their possible target genes in another independent sample by the methods of bioinformatics and Q-PCR. Thirdly, we will verify the target genes of these miRNAs, and further to determine these target genes and the upstream regulatory mechanism by reported genes and ChIP. Finally, we will illuminate the molecular mechanism of the miRNAs` and their target genes` effects on Glu-NMDAR neurotransmitter system and ΔfosB using methamphemine sensitizition mice model to up- or down- regulate the expression of the target genes in vivo by RNAi. The research will expand the molecular mechanism of methamphetamine addiction and provide new clues and ideas.