中期阿尔兹海默病presenilin-1/presenilin-2双基因敲除小鼠海马Vps33b基因的甲基化改变

Variation of Vps33b gene methylation in hippocampus of presenilin-1 / presenilin-2conditional double knockout mice at the mid-term of Alzheimer's disease

目的:探讨中期阿尔兹海默病的presenilin-1/presenilin-2双基因敲除小鼠(dK O小鼠)模型海马中Vps33b基因甲基化改变情况。方法:采用简化的表观亚硫酸盐测序技术(RRBS)检测12月龄雌性dK O小鼠和同龄雌性野生型小鼠各3只海马组织基因组DNA异常甲基化改变情况,利用相关生物软件对照分析获取异常甲基化基因,并通过重亚硫酸盐甲基化测序技术进行相应验证。结果:RRBS法筛选出Vps33b基因在中期阿尔兹海默病dK O小鼠海马中呈高甲基化状态,经重亚硫酸盐单基因测序验证Vps33b基因为高甲基化,与RRBS检测结果一致。结论:Vps33b基因在中期AD dK O小鼠海马中呈高甲基化状态,提示高甲基化的Vps33b基因可能与中期阿尔茨海默病病程相关。

Objective： The study was conducted to investigate the variation of Vps33 b gene methylation in hippocampus of presenilin-1 / presenilin-2 conditional double knockout mice（ dK O mice） at the mid-stage of Alzheimer＇s disease（ AD）. Methods： To identify the distribution and types of aberrant methylated genes,we detected genomic DNA in hippocampus of three female dK O mice and three female wild-type controls at 12 months of age with a method of reduced representation bisulphite sequencing（ RRBS）,compared the reference genome sequence with related biological software,and then verified by using single gene methylated sequencing. Results： The analysis by RRBS revealed hypermethylated Vps33 b gene existed in hippocampus of dK O mice at the mid-stage of neurodegeneration,and was confirmed by the result of single gene methylated sequencing. Conclusion： Hypermethylation of Vps33 b gene may be associated with the course of mid-term of AD in the dK O mice.