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目的探讨轴突导向分子EphA5在胚胎中脑神经上皮干细胞向靶组织纹状体特异性投射中的作用。方法自孕11d大鼠取材胚胎中脑神经上皮干细胞并进行体外培养,应用免疫细胞化学的方法检测EphA5在神经上皮干细胞中的表达情况;取材鼠脑纹状体组织,应用冰冻切片、RT-PCR及Western-blot等方法检测EphA5的配体Ephrin-A5的特异表达情况;将胚胎中脑神经上皮干细胞与新生鼠纹状体组织块联合培养,观察细胞的靶向生长情况。结果体外培养的胚胎中脑神经上皮干细胞可明显向纹状体生长并呈EphA5阳性,靶区纹状体组织中可检测到其配体Ephrin-A5高表达。结论轴突导向分子EphA5受体及其配体Ephrin-A5可介导胚胎中脑神经上皮干细胞向纹状体组织的靶向生长。
Objective To investigate the role of axon guidance molecule EphA5 in the specific projection of embryonic mesencephalic neural stem cells into the striatum of target tissues. Methods The embryonic midbrain neural stem cells were harvested from the 11th day of pregnancy and cultured in vitro. The expression of EphA5 in neuroepithelial stem cells was detected by immunocytochemistry. The striatum tissues were harvested and frozen sections were used for RT-PCR And Western-blot were used to detect the specific expression of EphA5 ligand Ephrin-A5. The embryonic mesencephalic neural stem cells were co-cultured with neonatal rat striatum to observe the target cell growth. Results Embryonic mesencephalic neural stem cells (SEGs) cultured in vitro could express striatum and showed EphA5-positive. Ephrin-A5 ligand was highly expressed in the target striatum. Conclusion Axonal targeting molecule EphA5 receptor and its ligand Ephrin-A5 can mediate the targeted growth of embryonic mesencephalic neural epithelial stem cells into striatum.