The effect of laminin-1 on enteric neural crest-derived cell migration in the Hirschsprung's disease mouse model
The effect of laminin-1 on enteric neural crest-derived cell migration in the Hirschsprung's disease mouse model
复制标题
Laminin-1对先天性巨结肠小鼠模型肠神经嵴源性细胞迁移的影响
DOI：
10.1007/s00383-017-4181-5
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发表时间：
2018
期刊：
影响因子：
1.8
通讯作者：
Yamataka A
中科院分区：
文献类型：
--
作者：
Nakazawa-Tanaka N;Fujiwara N;Miyahara K;Nakada S;Arikawa-Hirasawa E;Akazawa C;Yamataka A
层粘连蛋白-1调节多种神经细胞的突起生长。我们以前已经证明，层粘连蛋白-1促进肠神经嵴衍生细胞（ENCC）迁移，通过使用Sox 10-VENUS转基因小鼠，其中ENCC标记有绿色荧光蛋白，金星。缺乏内皮素-B受体基因的小鼠，Ednrb−/−小鼠，被广泛用作先天性巨结肠症（HD）的模型。本研究的目的是探讨层粘连蛋白-1（laminin-1）对新生HD小鼠（Sox 10-VENUS+/Ednrb−/−mice）ENCC迁移的影响。将标本与层粘连蛋白-1或不与层粘连蛋白-1孵育24小时，并在立体显微镜下拍摄图像。测量从胃到ENCC迁移波前的长度（L-E）和肠道总长度（L-G）。结果E12.5时，HD肠道样本中ENCC迁移的波前位于中肠，而Sox 10-VENUS+/Ednrb+/+（WT）样本中ENCC的波前已到达后肠。24小时后，HD肠道中的L-E/L-G从34.97%增加到36.46%，增加了1.49%，在含层粘连蛋白-1的HD中从48.08%增加到49.15%，提示层粘连蛋白-1给药对HD中ENCC迁移没有积极作用。1对E12.5 HD小鼠的ENCC迁移没有积极影响，与正常小鼠肠道标本中观察到的现象相反，其中层粘连蛋白-1在同一时期促进ENCC迁移。这表明ENCC与肠神经系统正常发育所需的细胞外环境因子之间的相互作用受损，导致HD中的无神经节结肠。
Background/aimLaminin-1 regulates neurite outgrowth in various neuronal cells. We have previously demonstrated that laminin-1 promotes enteric neural crest-derived cell (ENCC) migration by usingSox10-VENUS transgenic mice, in which ENCCs are labeled with a green fluorescent protein, Venus. Mice lacking the endothelin-B receptor gene,Ednrb−/−mice, are widely used as a model for Hirschsprung’s disease (HD). The aim of this study was to investigate the effects of laminin-1on ENCC migration inSox10-VENUS+/Ednrb−/−mice, a newly created HD mice model.MethodsFetal guts were dissected on embryonic day 12.5 (E12.5). Specimens were incubated either with, or without laminin-1 for 24 h and images were taken under a stereoscopic microscope. The length from the stomach to the wavefront of ENCC migration (L-E) and the total length of the gut (L-G) were measured. Changes in the ratio of L-E to L-G (L-E/L-G) after 24 h were calculated.ResultsOn E12.5, the wavefront of ENCC migration in the HD gut samples was located in the midgut, whereas the wavefront of ENCC inSox10-VENUS+/Ednrb+/+(WT) samples had reached the hindgut. After 24 h, L-E/L-G had increased by 1.49%, from 34.97 to 36.46%, in HD gut and had increased by 1.07%, from 48.08 to 49.15%, in HD with laminin-1, suggesting there was no positive effect of laminin-1 administration on ENCC migration in HD.ConclusionsOur results suggest that laminin-1 does not have a positive effect on ENCC migration in HD mice on E12.5, in contrast to the phenomenon seen in normal mice gut specimens, where laminin-1 promotes ENCC migration during the same period. This suggests that there is an impairment in the interaction between ENCC and extracellular environmental factors, which are required for normal development of the enteric nervous system, resulting in an aganglionic colon in HD.