PP1γ基因DNA甲基化在学习记忆中的作用  

Effect of expression of PP1γ and DNA methylation on learning and memory in mice

作　　者：侯林[1,2] 许晟迪 张柱霞 田小莉[1,2] 姜树原[1,2] 刘友[1,2] 巴德仁贵[1,2] 黄丽华[1,2] 邵国[1,2,3] 

机构地区：[1]包头医学院中心实验室生物医学研究中心基础学院包头医学院神经科学研究所,内蒙古包头014010 [2]内蒙古自治区低氧转化医学重点实验室,内蒙古包头014010 [3]首都医科大学宣武医院低氧适应转化医学北京重点实验室,北京100053

出　　处：《中国实验动物学报》2017年第2期146-152,共7页Acta Laboratorium Animalis Scientia Sinica

基　　金：国家自然科学基金(Nos:81460283;81660307);内蒙古自然科学基金(No:2014MS0810)

摘　　要：目的研究蛋白磷酸酶1γ(protein serine/threonine phosphatase,PP1γ)和DNA甲基化在学习记忆中的作用。方法通过DNA甲基化转移酶(DNA methytransferases,DNMTs)抑制剂5-aza-cdR处理小鼠,观察小鼠学习记忆情况及其PP1γ表达变化,通过水迷宫测定小鼠的学习记忆能力,Real-time PCR检测小鼠海马区DNMTs和PP1γmRNA转录水平以及Western-Blot测定PP1γ的蛋白质表达水平。为了进一步探讨5-aza-cdR对小鼠学习记忆的影响是否与细胞增殖和凋亡有关,用5-aza-cdR处理NG108-15神经细胞,流式细胞仪、x Celligence系统和荧光素酶报告基因分别检测5-aza-cdR对细胞增殖、凋亡和PP1γ的转录活性的影响。结果侧脑室注射了5-aza-cdR的小鼠空间学习记忆能力增加,同时小鼠海马区的DNMTs和PP1γ的表达降低;10μmol/L 5-aza-cdR抑制细胞增殖,降低PP1γ的转录活性,但是没有诱导细胞凋亡。结论 5-aza-cdR对PP1γ的表达抑制与小鼠学习记忆有关。Objectives To study the role of protein serine/threonine phosphatase1γ （PP1γ） and DNA methylation in learning and memory. Methods The mice and cells were treated with 5-aza-cdR, a DNA methyltransferase （DNMT） inhibitor, and to observe the changes of ability of learning and memory and expression levels of PP1γ in mice. The ability of learning and memory in mice was assessed by Morris Water maze test. The mRNA expression levels of DNMTs and PP1γ in the mouse hippocampus were determined by real-time PCR and the protein level of PP1γ was measured by Western blot. To further investigate their role, NG108-15 cell line was treated with 5-aza-cdR. Flow cytometry, xCelligence and luciferase were used to detect the effects of 5-aza-cdR on the proliferation, apoptosis and PP1γ transcription in the NG108-15 cells. Results The ability of learning and memory was enhanced in the mice after administration of 5-aza-cdR injection. The expressions of DNMTs and PP1γ were decreased in the mouse hippocampus after injection of 5-aza-cdR. On the other side, 10 μmol/L 5-aza-cdR inhibited cell proliferation and decreased PP1γ transcription without inducing apoptosis. Conclusions Our data demonstrate that 5-aza-cdR inhibits the expression of PP1γ which is related to learning and memory in mice.

关 键 词：蛋白磷酸酶1γ 5-AZA-CDR 海马 学习记忆 

分 类 号：Q95-33[生物学—动物学]