Clip170和Clasp1在小鼠神经系统中的表达及在自噬中作用  

The expression of Clip170 and Clasp1 in mouse nervous system and the role in autophagy

作　　者：于佳 甄然 彭宇 丁思妍 杨璇 宋彬彬 Yu Jia;Zhen Ran;Peng Yu;Ding Siyan;Yang Xuan;Song Binbin(Institute of Geriatrics and.Rehabilitation,the Beijing Geriatric Hospital,Beijing University of Chinese Medicine,Beijing 100095,China)

机构地区：[1]北京中医药大学附属北京老年医院老年医学基础研究中心,北京100095

出　　处：《脑与神经疾病杂志》2021年第10期649-652,共4页Journal of Brain and Nervous Diseases

基　　金：国家自然科学基金资助项目(82071438);北京市自然科学基金资助项目(7202077);北京市百千万人才工程资助项目(2017002);北京老年医院科研专项(2019bjlnyy-青-4)。

摘　　要：目的探讨微管正端示踪蛋白Clip170和Clasp1在小鼠神经系统中表达分布以及在细胞自噬蛋白降解通路中的作用。方法以1.5个月龄C57BL/6J野生型小鼠为材料,使用蛋白印迹(Western blot)检测Clip170和Clasp1在小鼠眼、脑、脊髓和坐骨神经中表达情况,以HEK293细胞为材料,使用转染和Western blot法分别检测HEK293细胞中Clip170和Clasp1缺失对于自噬通路中关键蛋白的影响。结果Clip170和Clasp1在小鼠神经系统中表达广泛,且与脊髓相比,坐骨神经中两者含量均显著下降(P<0.05)。在HEK293细胞中敲减Clip170蛋白可引起使自噬通路中p62和LC3B-Ⅱ蛋白表达水平显著提高(P<0.05),而LC3B-Ⅰ蛋白含量未发生明显变化(P>0.05)。HEK293细胞中敲减Clasp1蛋白可引起使自噬通路中p62蛋白表达水平极显著下降(P<0.05),LC3B-Ⅱ蛋白显著提高,LC3B-Ⅰ蛋白含量未发生明显变化(P>0.05)。结论微管正端示踪蛋白Clip170和Clasp1在小鼠神经系统广泛表达,但具有组织区域差异性。Clip170和Clasp1含量下降可能通过破坏自噬小体的形成或运输引起自噬通路异常。Objective To investigate the expression and distribution of microtubule plus-end tracking proteins Clip170 and Claspl in the nervous system in mice,and their role in autophagy.Methods The Clip170 and Claspl distribution in eye,olfactory bulb,cortex,hippocampus,striatum,midbrain,cerebellum,brainstem,spinal cord,sciatic nerve and muscle,and the expression of Clip170 and Claspl in spinal cord and sciatic nerve of 1.5 months wild-type mouse were examined by western blot.The Clip170 and Claspl function in autophagy of HEK293 cell were examined by siRNA transfection and western blot.Results Clip170 and Claspl had different expression levels in eye,brain,spinal cord,sciatic nerve and muscle.Compared to spinal cord,the sciatic nerve showed decreased level of Clip170 and Claspl(P<0.05).Knock down Clip170 of HEK293 cell increased the level of p62 and LC3 B-II(P<0.05).Knock down Claspl of HEK293 cell increased the level of LC3 B-I(P<0.05),but decreased the level of p62(P<0.05).And Both showed comparable level of LC3 B-II(P>0.05).Conclusion Clip170 and Claspl are distributed differently in the eyes,nervous system and muscles of mice.And decrease the level of Clip170 and Clasp1 can induce autophagy abnormal by regulating the formation or transport of autophagosomes.

关 键 词：Clip170 Clasp1 微管 自噬 

分 类 号：R744.8[医药卫生—神经病学与精神病学]