Sonic Hedgehog过表达下调后脑五羟色胺能神经元数量  

Sonic Hedgehog Overexpression Down-Regulates 5-hydroxytryptamine Neurons Number in Hindbrain

作　　者：谢洁 赵腾 贺白[2] XIE Jie;ZHAO Teng;HE Bai(Institute o f Neuroscience,Soochow University,Suzhou,Jiangsu,215123,China;Hematology department,The Third Affiliated Hospital of Soochow University,Changzhou,Jiangsu,213003,China)

机构地区：[1]苏州大学神经科学研究所,江苏苏州215123 [2]苏州大学附属第三医院血液科,江苏常州213003

出　　处：《现代生物医学进展》2018年第10期1870-1874,共5页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(81330026)

摘　　要：目的:本研究主要是探索高浓度的Shh对后脑5-HT神经元数量的影响。方法:通过免疫荧光和原位杂交手段检测Shh在脑干的表达情况。离体培养5-HT神经元,用不同浓度Shh蛋白处理,观察5-HT神经元的数量变化以及对轴突的影响。通过胚胎宫内电转,检测Shh过表达后脑5-HT神经元的数量变化。结果:Shh在脑干5-HT神经元分布区域内表达。离体培养的5-HT神经元,250 ng/m L的Shh蛋白处理后神经元数量为41.25±0.52(n=4,P=0.0218),与对照组35±1.21(n=4)相比,神经元数量上调。相反,1250 ng/m L的Shh蛋白处理后神经元数量为7.5±0.43(n=4,P<0.0001),与对照组相比,神经元数量极显著下降。250 ng/m L的Shh蛋白处理后5-HT神经元轴突长度为1.08±0.05(n=4,P=0.7555),与对照组1±0.01(n=4)相比,轴突长度没有显著性差异。然而1250 ng/m L的Shh蛋白处理后5-HT神经元轴突长度为0.44±0.03(n=4,P=0.0014),与对照组相比,轴突长度极显著缩短。胚胎宫内电转p IRES-Shh-EGFP和p IRES-EGFP,观察到Shh过表达缝核上行5-HT神经元数量为147±54.2(n=4,P=0.0053),相较于对照组459±49.0(n=4),神经元数量极显著下降。同样地,Shh过表达缝核下行5-HT神经元数量为187±18.4(n=4,P=0.0001),相较于对照组411±17.3(n=4),神经元数量也发生了极显著下降。结论:Shh过表达对5-HT神经元的发育有负向的调控作用,主要表现在引起后脑缝核5-HT神经元数量减少。Objective： The aim of this study was to explore the effect of high concentration of Sonic Hedgehog（Shh） on 5-hydroxytryptamine（5-HT） neurons number in hindbrain.Methods： Immunofluorescence and in situ hybridization were performed to detect the expression of Shh in brainstem.Cultured 5-HT neurons were treated with different Shh gradients,to observe the change of neurons number and effect on axons.The change of 5-HT neurons number was detected through Shh overexpression by in utero electroporation.Results：Shh was expressed in brainstem where 5-HT neurons distributed.In vitro cultured 5-HT neurons,treated with 250 ng/m L Shh protein,neurons number was 41.25±0.52（n=4,P=0.0218）,and comparing to control groups 35±1.21（n=4）,there was an increase of neurons number.In contrast,treated with 1250 ng/m L Shh protein,neurons number was 7.5±0.43（n=4,P〈0.0001）,and there was an significant reduction of neurons number comparing to control groups.Besides,treated with 250 ng/ml Shh protein,the length of 5-HT axons was1.08±0.05（n=4,P=0.7555）,and comparing to control groups 1±0.01（n=4）,there was no detectable effect on the axon length.However,treated with 1250 ng/m L Shh protein,the length of 5-HT axons was 0.44±0.03（n=4,P=0.0014）,if compared to control groups,the axon length was remarkably shortened.In utero electroporation with p IRES-Shh-EGFP and p IRES-EGFP,observed the number of ascending5-HT neurons in raphe nuclei was 147±54.2（n=4,P=0.0053） through Shh overexpression,and neurons number decreased notably comparing to control groups 459±49.0（n=4）.Similarly,the number of 5-HT neurons in raphe nuclei was 187±18.4（n=4,P=0.0001） through Shh overexpression,and comparing to control groups 411±17.3（n=4）,neurons number remarkably reduced.Conclusion： Shh overexpression negatively regulated the development of 5-HT neurons,and mainly reflected in leading to the reduction of 5-HT neurons in raphe nuclei of hindbrain.

关 键 词：SHH 5-HT神经元 过表达 

分 类 号：R-332[医药卫生] Q593.2[生物学—生物化学]