PD相关基因LRRK2表达调控的初探

Expression and regulation research of PD-related gene LRRK2

目的:以C57BL/6J(B6)和DBA/2(D2)小鼠为实验动物,初步探究帕金森病(PD)相关基因LRRK2的表达调控机制,构建LRRK2的表达调控网络。方法:数量性状基因座(eQTL)分析技术分析LRRK2基因在各品系小鼠中的表达情况,并进行遗传学相关分析和区间连锁分析;半定量PCR技术和Real-time PCR检测LRRK2基因在PD模型组和对照组小鼠中的表达量。结果:与NS对照组相比,B6、D2小鼠PD模型组的中脑及前脑中LRRK2基因的mRNA表达水平都发生了非常显著的变化(P<0.01)。eQTL分析发现在Affymetrix MOE430芯片中,LRRK2基因共有2个探针位置用于分析其在前脑、中脑的mRNA表达水平。选取探针位置1431394_a_at_A进行检测,结果显示,LRRK2基因在亲本B6、D2及41个BXD RI小鼠前脑、中脑的表达在各品系间的表达量多少不一。遗传学相关分析结果揭示,LRRK2基因与100个基因高度相关,初步构建LRRK2基因的调控网络,该网络包括了Kif21a等21个与LRRK2基因高度相关的基因。区间连锁分析检测引起探针位置1431394_a_at_A表达水平差异的染色体区域,结果显示在15号染色体和4号染色体各存在一个LRS≥20的QTL峰。结论:LRRK2基因既存在顺式调控机制又存在反式调控机制。

Objective：To study initially the expression regulation mechanism and regulatory networks for LRRK2 gene related PD using C57BL/6J（B6） and DBA/2（D2） mouse as experimental animals.Methods：Using eQTL analysis technology,Genetics-related analysis as well as interval linkage analysis to detect the LRRK2 gene expression value in mouse;and semi-quantitative PCR was used to detect the expression of LRRK2 gene in PD model group and control group.Results：Compared with the NS control group,the expression of LRRK2 gene mRNA have undergone significant changes in the midbrain and forebrain of PD model group（P0.01）.The eQTL analysis for the LRRK2 gene revealed that there were the two Probe Sets in the Affymetrix MOE430 chip,which could be used to analyze the mRNA expression levels.We select Probe Set 1431394_a_at_A for subsequent analysis due to its high expression and because it is located in the encoding area.The eQTL detection revealed that the LRRK2 expression levels were different between the two different strains.Genetics-related analysis proved that there were one hundred genes highly related to LRRK2 gene.We initially constructed the LRRK2 gene regulatory network,which included twenty-one genes highly related with LRRK2 such as Kif21a.The interval linkage analysis showned that the chromosomal regions caused the differences in expression levels of Probe Set 1431394_a_at_A,resulted in two peaks with LRS（Likelihood ratio statistics,the level of chain groups significant threshold） ≥20 at chromosome 15 and chromosome4.Conclusion：LRRK2 gene expression regulation exits both cis-regulatory and trans-regulatory mechanism.