背侧抑制性轴突导向蛋白对胎鼠脊髓神经嵴细胞迁移特性的影响

崔祖旗  缪晓锦  谷泽林  巩梦飞  陈欢  杨舒涵 刘桐妤  张三兵 苏玉红

doi:10.16098/j.issn.0529-1356.2025.02.004

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解剖学报 ›› 2025, Vol. 56 ›› Issue (2) : 150-157. DOI: 10.16098/j.issn.0529-1356.2025.02.004

神经生物学

背侧抑制性轴突导向蛋白对胎鼠脊髓神经嵴细胞迁移特性的影响

崔祖旗¹ 缪晓锦² 谷泽林² 巩梦飞² 陈欢² 杨舒涵² 刘桐妤² 张三兵^3* 苏玉红^2*

作者信息 +

Effect of Draxin on the migration characteristics of trunk neural crest cell in the embryonic mouse spinal cord

CUI Zuqi¹ MIAO Xiao-jin² GU Ze-lin² GONG Meng-fei² CHEN Huan² YANG Shu-han² LIU Tong-yu² ZHANG San-bing^3* SU Yu-hong^2*

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摘要

目的探讨胎鼠脊髓早期发育过程中背侧抑制性轴突导向蛋白（Draxin）对神经嵴细胞迁移特性的影响。方法应用免疫组织化学、原位杂交等方法，检测Draxin在发育早期胎鼠脊髓内的表达特性(每组8只小鼠)；应用原位杂交的方法，检测发育早期不同类型的胎鼠脊髓内神经嵴细胞迁移特性的改变(每组5只小鼠)；应用体外培养的方法，观察Draxin对体外培养的神经嵴细胞迁移特性的影响(每组16只小鼠)。结果敲除Draxin基因的同时引入β-半乳糖苷酶基因Z（LacZ）从而制备相关基因敲除小鼠，应用β-半乳糖苷酶染色方法检测Draxin敲除胎鼠体内LacZ基因表达情况，结果显示，胚胎9.5 d(E9.5）时期开始胎鼠脊髓内可见Draxin表达，E10.5时期胎鼠脊髓内Draxin表达明显，应用原位杂交方法检测野生型胎鼠脊髓内Draxin基因表达情况，结果进一步验证E10.5时期胎鼠脊髓内Draxin表达明显；应用Sox10反义探针原位杂交的方法检测胎鼠脊髓内神经嵴细胞迁移情况，结果显示，E9.5~E10.5为胎鼠脊髓内神经嵴细胞大量迁移的时期，且与野生型小鼠相比，部分Draxin敲除导致胎鼠脊髓内神经嵴细胞节段性迁移特性的形成延迟；应用体外培养的方法检测Draxin对体外培养的野生型胎鼠脊髓神经嵴细胞迁移特性的影响，结果显示，Draxin使体外培养的神经嵴细胞迁移距离下降。结论胎鼠发育早期（E9.5~E10.5），脊髓内神经嵴细胞迁移旺盛，同一时期Draxin通过排斥性调控作用引起神经嵴细胞远离表达Draxin的区域，从而对神经嵴细胞特异性迁移路径的形成起重要的调控作用。

Abstract

Objective To investigate the effect of dorsal repulsive axon guidance protein（Draxin）on the migration of trunk neural crest cells during the early development of embryonic mouse spinal cord. Methods Immunohistochemistry and in situ hybridization were used to detect the expression characteristics of Draxin in early embryonic spinal cord (8 mice each group); In situ hybridization was used to detect the change of migration characteristics of trunk neural crest cells in early embryonic spinal cord of different types of mouse (5 mice each group); in vitro culture method was used to check the effect of Draxin on the migration characteristics of embryonic mouse trunk neural crest cells (16 mice each group). Results β-galactosidase gene Z(LacZ) gene was introduced when Draxin gene was knocked out to produce Draxin gene knockout mice. β-galactosidase staining was used to detect LacZ gene expression in Draxin knockout embryonic mice, and the result showed that Draxin expression was observed in the spinal cord of early embryonic mice since 9.5 days(E9.5). Draxin expression was obvious in the embryonic mice spinal cord in E10.5 period. In situ hybridization was used to detect the expression of Draxin gene in the spinal cord of wild type embryonic mice, and the result further verified the obvious expression of Draxin in the early embryonic mice spinal cord in E10.5 period. Sox10 in situ hybridization was used to detect neural crest cell migration in the spinal cord of embryonic mice in E10.5 period. The result showed that segmental migration of neural crest cells in the early embryonic spinal cord of some Draxin knockout mice was delayed compared with the wild type mice. The effect of Draxin on the migration of wild type early embryonic mice trunk neural crest cells in vitro was tested. The result showed that Draxin reduced the migration distance of neural crest cells in vitro. Conclusion In the early developmental stage of embryonic spinal cord (E9.5-E10.5), neural crest cells migrated exuberant. At the same time, Draxin plays an important inhibitory function in the formation of the specific migration pathways of trunk neural crest cells by promoting neural crest cells migrating away from Draxin expressing regions.

导出引用

崔祖旗缪晓锦谷泽林巩梦飞陈欢杨舒涵刘桐妤张三兵苏玉红. 背侧抑制性轴突导向蛋白对胎鼠脊髓神经嵴细胞迁移特性的影响[J]. 解剖学报. 2025, 56(2): 150-157 https://doi.org/10.16098/j.issn.0529-1356.2025.02.004

CUI Zuqi MIAO Xiao-jin GU Ze-lin GONG Meng-fei CHEN Huan YANG Shu-han LIU Tong-yu ZHANG San-bing SU Yu-hong. Effect of Draxin on the migration characteristics of trunk neural crest cell in the embryonic mouse spinal cord[J]. Acta Anatomica Sinica. 2025, 56(2): 150-157 https://doi.org/10.16098/j.issn.0529-1356.2025.02.004

中图分类号： R394.1

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