鸡胚发育过程中Shh超表达对脊髓神经纤维投射的抑制作用

Spinal cord nerve fiber projection is repressed by sonic hedgehog overexpression during chicken embryonic development

目的探讨鸡胚发育过程中,sonic hedgehog（Shh）在脊髓中超表达后对神经纤维投射的影响。方法在鸡胚龄2.5～3d（E2.5～E3）时,利用鸡胚活体原位电转基因技术将Shh表达质粒和携带有报告基因绿色荧光蛋白（GFP）的质粒共转入鸡胚脊髓,E6时取材切片,至少取3个材料,免疫荧光技术检验Shh的超表达,利用Open Book技术研究Shh在鸡胚脊髓中超表达后对神经纤维投射的影响。结果在脊髓组织切片水平上,免疫荧光结果表明,Shh在鸡胚脊髓中成功超表达,无论在组织切片上还是通过Open Book技术进行观察,与对照组相比,超表达后转染侧神经纤维都无法正常通过底板投射到对侧,表明Shh在鸡胚脊髓神经纤维投射方面具有重要作用。结论在鸡胚发育过程中,脊髓中Shh超表达后对神经纤维投射具有抑制作用。

Objective To study the effect of sonic hedgehog（Shh） overexpression on nerve fiber projection in the spinal cord during chicken embryonic development. Methods At embryonic day （E）2. 5-3.0, in vivo electroporation was used to co-transfect Shh expression plasmid and reporter gene plasmid into the chicken embryonic spinal cord. Samples were collected and sliced at E6. At least three samples were collected. The overexpression of Shh was detected using immunofluorescence histochemistry, and nerve fiber projecting affected by Shh overexpression was detected with Open Book technology. Results The results of immunofluorcscence showed Shh was overcxpressed in the chicken embryonic spinal cord successfully. Compared with the control group, after Shh overexpression, the nerve fiber did not across the floor plate and project stereotypically both in slice level and whole mount level, which suggested that Shh plays an important role for nerve fiber projecting in the chicken embryonic spinal cord. Conclusion The normal nerve fiber projection is repressed by Shh overexpression in the chicken embryonic spinal cord.