| 谢 洁,赵 腾,贺 白.Sonic Hedgehog过表达下调后脑五羟色胺能神经元数量[J].现代生物医学进展英文版,2018,(10):1870-1874. |
| 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 |
| Received:June 30, 2017 Revised:July 25, 2017 |
| DOI：10.13241/j.cnki.pmb.2018.10.013 |
| 中文关键词: Shh 5-HT神经元 过表达 |
| 英文关键词: Shh 5-HT neurons Overexpression |
| 基金项目:国家自然科学基金项目（81330026） |
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| 中文摘要: |
| 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/mL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/mL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/mL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/mL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pIRES-Shh-EGFP和pIRES-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。 |
| 英文摘要: |
| ABSTRACT Objective: The aim of this study was to explore the effect of high concentration of Sonic Hedgehog (Shh) on 5-hydrox- ytryptamine (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/mL 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/mL 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 was 1.08±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/mL 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 pIRES-Shh-EGFP and pIRES-EGFP, observed the number of ascending 5-HT neurons in raphe nuclei was 147±54.2(n=4, P=0.0053) through Shh overexpression, and neurons number decreased notably com- paring 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 overex- pression 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. |
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