母婴分离应激对幼年大鼠脑衰反应调节蛋白2及微管动态性影响  

Effects of early maternal separation on CRMP2 and microtubule dynamics in the hippocampus of young rats

作　　者：魏艳艳 陈景旭[1] 吴作天 肖玲[2] 何静 张楠 王高华[2] WEI Yanyan;CHEN Jingxu;WU Zuotian;XIAO Ling;HE Jing;ZHANG Nan;WANG Gaohua(Renmin Hospital of Wuhan University,Wuhan 430060,China)

机构地区：[1]北京回龙观医院 [2]武汉大学人民医院精神卫生中心,武汉430060

出　　处：《中国神经精神疾病杂志》2020年第7期400-404,共5页Chinese Journal of Nervous and Mental Diseases

基　　金：国家自然科学基金资助项目(编号:81571325,81871072);首发专项基金资助项目(编号:2018-3-2132)。

摘　　要：目的探讨不同程度母婴分离(maternal separation,MS)应激对幼年雄性大鼠海马脑衰反应调节蛋白2(collapsin response mediator protein 2,CRMP2)及微管动态性的影响。方法36只新生雄性大鼠随机分为3组,母婴分离360 min组(MS360组)、母婴分离15 min组(MS15组)及对照组(NC组)。出生后第4~10天进行1周的母婴分离,每天分离360 min或15 min,NC组正常饲养。采用q-RT-PCR技术检测海马CRMP2、actin、tubulin的mRNA表达水平,采用Western-Blot技术检测海马CRMP2、P-CRMP2、actin、tubulin及动态微管标志物Tyr-tubulin、稳定微管标志物Acet-tubulin表达水平。结果3组海马P-CRMP2、Acet-tubulin、Tyr-tubulin表达水平差异有统计学意义(P<0.05),MS360组与MS15组及NC组大鼠相比,P-CRMP2、Acet-tubulin表达水平升高(P<0.05),Tyr-tubulin表达水平下降(P<0.05),MS15组与NC组无统计学差异(P>0.05)。3组CRMP2、actin、tubulin的mRNA和蛋白表达水平无统计学差异(P>0.05)。结论长时间母婴分离应激可升高CRMP2的磷酸化水平,降低海马微管动态性,使神经可塑性受损,而短期母婴分离未对海马微管动态性产生不利影响。Objective To investigate the effects of different degrees of maternal separation(MS)stress on hippocampus CRMP2,P-CRMP2 expression and microtubule dynamics in young male rats.Methods Thirty-six newborn male rats were randomly divided into three groups:maternal separation 360 min group(MS360 group),maternal separation 15 min group(MS15 group)and normal control group(NC group).The maternal separation was performed for 1 week from postnatal day(PND)4 to PND10 for 360 min or 15 min per day.The NC group was reared normally.Q-RT-PCR was used to detect the mRNA expression levels of CRMP2,actin and tubulin.Western-Blot technology was used to detect the expression levels of CRMP2,P-CRMP2,actin,tubulin,dynamic microtubule marker Tyr-tubulin and stable microtubule marker Acet-tubulin.Results The expression levels of P-CRMP2,Acet-tubulin and Tyr-tubulin in the hippocampus were significantly different among the three groups(P<0.05).Compared with MS15 group and the NC group,the expression levels of P-CRMP2 and Acet-tubulin were higher(P<0.05),and the expression level of Tyr-tubulin was lower(P<0.05)in MS360 group.The mRNA and protein expression levels of CRMP2,actin and tubulin were not significantly different among the three groups(P>0.05).Conclusion Prolonged maternal separation stress can significantly increase CRMP2 phosphorylation levels,reduce hippocampal microtubule dynamics,and impair neuroplasticity.Brief maternal separation has no adverse effect on hippocampal microtubule dynamics.

关 键 词：母婴分离 脑衰反应调节蛋白2 微管动态性 神经可塑性 

分 类 号：R74[医药卫生—神经病学与精神病学]