背侧抑制性轴突导向蛋白的敲减对鸡胚后脑内23C10阳性神经纤维投射的影响

王滋怡; 张舒涵; 张子韧; 张文文; 胡亚男; 张三兵; 苏玉红

doi:10.16098/j.issn.0529-1356.2022.04.003

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解剖学报 ›› 2022, Vol. 53 ›› Issue (4) : 418-423. DOI: 10.16098/j.issn.0529-1356.2022.04.003

神经生物学

背侧抑制性轴突导向蛋白的敲减对鸡胚后脑内23C10阳性神经纤维投射的影响

王滋怡¹ 张舒涵² 张子韧² 张文文² 胡亚男² 张三兵^3* 苏玉红^2*

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Effect of draxin knockdown on the projection of 23C10-positive neural fibers in the embryonic chick hindbrain

WANG Zi-yi¹ ZHANG Shu-han² ZHANG Zi-ren² ZHANG Wen-wen² HU Ya-nan² ZHANG San-bing^3* SU Yu-hong^2*

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摘要

目的探讨敲减鸡胚后脑内背侧抑制性轴突导向蛋白（draxin）的表达对23C10阳性神经纤维投射特性的影响。方法以碱性磷酸酶（ALP）融合蛋白与HH21~22阶段鸡胚后脑活组织切片体外孵育为对照组，以draxin-ALP融合蛋白与HH21~22阶段鸡胚后脑活组织切片体外孵育为实验组，每组10例，检测鸡胚后脑内的23C10阳性神经纤维是否具有与draxin直接结合的能力；以鸡胚后脑内空白质粒电穿孔组为对照，以针对draxin表达的小干扰RNA（siRNA）表达载体质粒电穿孔为实验组，每组18例，观察敲减draxin的效果，以及是否引起鸡胚后脑内23C10阳性神经纤维投射特性的改变。结果 HH21~22阶段鸡胚后脑活组织切片内，大部分draxin阳性信号与23C10阳性神经纤维重叠；应用电穿孔方法导入draxin的siRNA敲减质粒后，draxin相对表达量显著降低（P<0.05）；HH25~26阶段的鸡胚后脑内，电穿孔一侧的23C10阳性神经纤维的背侧区域内投射纤维分布分散（P<0.05）。结论鸡胚后脑发育过程中，draxin可以与23C10阳性神经纤维直接结合，且其在鸡胚后脑内23C10阳性神经纤维成束投射的形成过程中发挥重要的调控作用。

Abstract

Objective To investigate the effects of the downregulation of draxin expression on the projection characteristics of 23C10-positive neural fibers in the chick embryonic hindbrain. Methods The vitro incubation of HH stages 21-22 chick embryonic hindbrain biopsy with alkaline phosphatase （ALP）protein was used as control group. The incubation of HH stages 21-22 chick embryonic hindbrain biopsy with draxin-ALP fusion protein was used as experimental group. The number of embryonic hindbrain for each group was 10. To detect whether 23C10-positive neural fibers could directly bind to draxin protein or not；In ovo electroporation using empty vector in the chick embryonic hindbrain was used as control group. In ovo electroporation with small interfering RNA(siRNA) expressing vector for reducing draxin expression in the chick embryonic hindbrain was used as experimental group. The number of embryonic hindbrain for each group was 18. The effect of the down-regulation of draxin expression and the change of projection characteristics of 23C10-positive neural fibers were observed to check whether the down-regulation of draxin expression would affect the distribution of 23C10-positive fibers. Results Most portion of draxin protein could overlap with 23C10-positive neural fibers in HH stages 21-22 chick embryonic hindbrain biopsies；After expression of the siRNA plasmid against draxin by electroporation, the expression level of draxin protein was significantly reduced, and the distribution of 23C10-positive fibers was scattered in the dorsal hindbrain on the electroporated side at HH stages 25-26 of chick embryos（P<0.05）. Conclusion Draxin protein may directly bind to 23C10-positive fibers in hindbrain, and it plays an important regulatory role in the fasciculation of 23C10-positive fibers during chick embryonic development.

导出引用

王滋怡张舒涵张子韧张文文胡亚男张三兵苏玉红. 背侧抑制性轴突导向蛋白的敲减对鸡胚后脑内23C10阳性神经纤维投射的影响[J]. 解剖学报. 2022, 53(4): 418-423 https://doi.org/10.16098/j.issn.0529-1356.2022.04.003

WANG Zi-yi ZHANG Shu-han ZHANG Zi-ren ZHANG Wen-wen HU Ya-nan ZHANG San-bing SU Yu-hong. Effect of draxin knockdown on the projection of 23C10-positive neural fibers in the embryonic chick hindbrain[J]. Acta Anatomica Sinica. 2022, 53(4): 418-423 https://doi.org/10.16098/j.issn.0529-1356.2022.04.003

中图分类号： R394.1

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