Spastin基因突变对斑马鱼脊髓运动神经元的作用研究  

Effects of Spastin mutation on the spinal motor neurons in zebrafish

作　　者：赖晓娜 李媛媛 陈旭 巫荣华 董张及 刘梅[1] LAI Xiaona;LI Yuanyuan;CHEN Xu;WU Ronghua;DONG Zhangji;LIU Mei(Key Laboratory of Neuroregeneration of Jiangsu and Ministry of Education,Co-innovation Center of Neuroregeneration,Nantong University,Nantong 226001)

机构地区：[1]南通大学教育部/江苏省神经再生重点实验室/神经再生协同创新中心,南通226001

出　　处：《南通大学学报（医学版）》2021年第1期1-5,共5页Journal of Nantong University(Medical sciences)

基　　金：国家自然科学基金资助项目(31701049);江苏省自然科学基金面上项目(BK20171253)。

摘　　要：目的:微管切割蛋白Spastin是遗传性痉挛性截瘫(hereditary spastic paraplegia,HSP)的主要致病基因,本文旨在探讨Spastin基因突变对运动神经系统的形态和功能的影响。方法:鉴定斑马鱼spost敲除突变体的纯合子和杂合子,检测包括野生型的3种基因型spest^(+/+)、spest^(+/ZKO988a)、spest^(ZKO988a/ZKO988a)斑马鱼在无刺激下、光刺激下及声音刺激下自由游泳的运动速率;激光共聚焦显微镜成像转基因spast敲除突变体斑马鱼3种基因型Tg(hb9:eCFP);spost、Tg(hb9:eCFP);spost^(+/ZKO988a)、Tg(hb9:eGFP);spast^(ZKO988a/ZKO988a)的脊髓运动神经元(Cap)形态差异。结果:突变体spest^(+/ZKO988a)和spest^(ZKO988a/ZKO988a)斑马鱼的运动速率与野生型无差异;突变体Tg(hb9:eGFP);spast^(+/ZKO988a)和Tg(hb9:eGFP);spast^(ZKO988a/ZKO988a)斑马鱼朝向腹侧的CaP脊髓运动神经元的轴突长度显著短于野生型的轴突长度。结论:Spastin M1蛋白缺失对斑马鱼的运动速率无显著改变,但导致CaP神经元轴突长度缩短。Objective:Spastin,a microtubule-severing protein,is the main pathogenic gene for hereditary spastic paraplegia(HSP).This article aims to investigate the effects of Spastin mutation on the functions of spinal motor neurons in zebrafish.Methods:The genotypes of spast^(+/+),spost^(+/ZKO988a),spast^(ZKO988a/ZKO988a)were identified,and their swimming velocity were recorded and analyzed under non-stimulation,light-stimulation and sound-stimulation respectively;the spinal motor neuron(CaP)neuronal phenotype of spast mutant zebrafish:Tg(hb9:eGFP);spast^(+/+),Tg(hb9:eGFP);spast^(+/ZKO988a),Tg(hb9:eGFP);spast^(ZKO988a/ZKO988a)were observed by confocal microscopy.Results:The swimming velocity of the spast^(+/ZKO988a)and spart(ZKO988a/ZKO988a)zebrafishes was no significant difference with spast wild type zebrafishes;the axonal length of CaP neurons of Tg(hb9:eGFP);spast^(+/ZKO988a)and Tg(hb9:eGFP);spast^(ZKO988a/ZKO988a)zebrafishes were significant reduced,compared with the spast wild type zebrafishes.Conclusion:Spastin M1 mutation resulted in no alteration on zebrafishes’locomotor function,but reduced the axonal length of CaP neurons.

关 键 词：spast 遗传性痉挛性截瘫 脊髓运动神经 斑马鱼 

分 类 号：Q78[生物学—分子生物学]