Nakazawa Nana, Miyahara Katsumi, Okawada Manabu, Yamataka Atsuyuki, Suzuki Ryota, Akazawa Chihiro, Tomikawa-Ichikawa Naoki, Arikawa-Hirasawa Eri
Department of Pediatric Surgery, Juntendo Nerima Hospital, 3-1-10 Takanodai, Nerima-ku, Tokyo, 177-8521, Japan,
Pediatr Surg Int. 2013 Nov;29(11):1205-8. doi: 10.1007/s00383-013-3388-3.
BACKGROUND/AIM: Neuronal development is regulated by extracellular environmental factors including nerve growth factor (NGF) and laminin. We have previously demonstrated that laminin-1 promotes neurite outgrowth of dorsal root ganglion cells by modulating NGF and integrin signaling. However, information about their effects on the enteric nervous system (ENS) is limited. Recently, we succeeded in visualizing enteric neural crest-derived cell (ENCC) migration using SOX10-Venus transgenic mice, in which ENCC are labeled with a green fluorescent protein, Venus. In this study, we examine the effects of NGF and laminin-1 in ENCC migration using SOX10-Venus mice gut.
Pregnant SOX10-Venus mice were killed on day 12.5 of gestation. The colorectum was dissected from embryos (n = 10) and placed in culture medium including NGF with or without laminin-1 for 12 h. Extension rates of ENCC migration, colorectum and ENCC migration per colorectum were calculated.
Venus positive-ENCC extension rate was significantly higher in the laminin group (n = 5) compared to control (n = 5), 22.84 and 13.96 %, respectively (p < 0.05). The extension rate of the colorectum was not significantly different between the two groups.
Our results suggest that laminin promotes ENCC migration in mice. This technique allowed us to visualize the effects of extracellular molecules on ENCC migration and it potentially provides us with an insight into the pathophysiology of developmental disorders of the ENS, such as Hirschsprung's disease.
背景/目的：神经元发育受包括神经生长因子（NGF）和层粘连蛋白在内的细胞外环境因素调控。我们之前已经证明，层粘连蛋白-1通过调节NGF和整合素信号通路促进背根神经节细胞的神经突生长。然而，关于它们对肠神经系统（ENS）影响的信息有限。最近，我们利用SOX10-维纳斯转基因小鼠成功观察到肠神经嵴衍生细胞（ENCC）的迁移，其中ENCC被绿色荧光蛋白维纳斯标记。在本研究中，我们使用SOX10-维纳斯小鼠肠道研究NGF和层粘连蛋白-1对ENCC迁移的影响。
妊娠12.5天的SOX10-维纳斯小鼠被处死。从胚胎（n = 10）中分离出结直肠，置于含有或不含有层粘连蛋白-1的NGF培养基中培养12小时。计算ENCC迁移的延伸率、结直肠和每结直肠的ENCC迁移率。
与对照组（n = 5）相比，层粘连蛋白组（n = 5）中维纳斯阳性ENCC的延伸率显著更高，分别为22.84%和13.96%（p < 0.05）。两组之间结直肠的延伸率没有显著差异。
我们的结果表明层粘连蛋白促进小鼠ENCC迁移。这项技术使我们能够观察细胞外分子对ENCC迁移的影响，并可能为我们深入了解ENS发育障碍（如先天性巨结肠）的病理生理学提供线索。
