目的探讨不同发育时期的鸡胚脊髓内背侧抑制性轴突导向蛋白(draxin)表达与神经嵴迁移之间在空间分布上的相关性。方法选取HH 15-16、HH19-20阶段鸡胚各10只,应用原位杂交、免疫组织化学等方法,观察各阶段鸡胚脊髓内draxin的表达部位及神经嵴细胞迁移路径,同时比较两者在空间分布上的相关性;应用活组织切片与碱性磷酸酶标记的draxin融合蛋白(draxin—AP)体外孵育的方法,检测迁移路径内的神经嵴细胞是否具有与draxin蛋白直接结合的能力。结果随着发育时间的推移,鸡胚脊髓内draxin的表达和神经嵴迁移都具有明显的由颅侧向尾侧渐进性分布的特性,且draxin的表达部位位于神经管背侧、顶板和背侧生皮肌节之间,这些部位恰均位于神经嵴迁移路径的周围;同时迁移路径内的神经嵴细胞具有与draxin蛋白体外直接结合的能力。结论draxin表达在神经嵴迁移路径周围且部分神经嵴具有与draxin蛋白体外直接结合的能力,推测draxin可能在鸡胚脊髓神经嵴迁移过程中发挥重要的调节作用。
Objective To examine the correlation of draxin expression and neural crest migration in chick embryonic spinal cord at different developmental stages. Methods Ten chick embryos at different stages were used. In situ hybridization and immunohistochemistry were used to observe the localization of draxin expression, the migration pathway of neural crest cell-the spatial correlation of draxin expression and the neural crest migration. Living tissue incubation with draxin alkaline phosphatase (draxin-AP) recombinant protein was used to check the binding ability of migrating neural crest with draxin protein. Results With development, the draxin expression and neural crest migration had a very similar anterior- posterior gradient in chick embryonic spinal cord. Draxin was expressed in the dorsal neural tube, roof plate and dorsal tip of dermomyotome which surrounded the migration pathway of neural crest cells. We also found that portion of migrating neural crest cells bound to draxin-AP protein directly. Conclusion Draxin is expressed in the tissues surrounding the neural crest migration pathway and portion of migrating neural crest cells binds to draxin protein directly. We conclude therefore that draxin may be involved in the regulation of neural crest cell migration in chick embryonic spinal cord.