Newly raised anti-c-Kit antibody visualizes morphology of interstitial cells of Cajal in the developing gut of chicken embryos.
Newly raised anti-c-Kit antibody visualizes morphology of interstitial cells of Cajal in the developing gut of chicken embryos.
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新产生的抗 c-Kit 抗体可观察鸡胚胎发育肠道中 Cajal 间质细胞的形态。
DOI：
10.1111/dgd.12808
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Y
中科院分区：
文献类型：
--
作者：
Yagasaki;R.;Shikaya;Y.;Kawachi;T.;Inaba;M.;Takase;Y.;Takahashi;Y
肠道蠕动运动是局部收缩的波状传播，对于摄入物质的运输和消化非常重要。在肠神经系统（ENS）、平滑肌细胞和卡哈尔间质细胞（ICC）这三种细胞中，ICC被认为起着起搏器的作用，因此破译ICC的细胞功能对于进一步了解肠道蠕动非常重要。 c-Kit 是一种酪氨酸激酶受体，已广泛用作 ICC 的标记物。大多数关于 ICC 的研究都是使用市售的抗 c-Kit 抗体在哺乳动物中进行的。最近，鸡胚肠道已成为研究肠道蠕动的强大模型。然而，由于针对哺乳动物的抗 c-Kit 抗体不适用于鸡，因此 ICC 调节的细胞机制在很大程度上尚未被探索。在这里，我们报道了一种新产生的针对鸡 c-Kit 蛋白的多克隆抗体。通过蛋白质印迹分析和免疫细胞化学验证了抗体的特异性。使用新型抗体和抗 α 平滑肌肌动蛋白 (αSMA) 抗体进行联合免疫染色，成功地观察了正在发育的鸡后肠的环肌和纵肌层中的 ICC。正如之前在小鼠中所显示的，早期的 ICC 和平滑肌细胞的共同祖细胞对 αSMA 和 c-Kit 双阳性，而在后期，分化的 ICC 和平滑肌细胞分别仅表现出 c-Kit 和 αSMA。还发现了一种新的 ICC 群体，从粘膜下层放射状延伸到环形肌层。此外，新抗体在清晰的后肠中描绘出了单个 ICC。本研究新开发的抗体将有助于鸡胚胎蠕动运动的研究。
The gut peristaltic movement, a wave‐like propagation of a local contraction, is important for the transportation and digestion of ingested materials. Among three types of cells, the enteric nervous system (ENS), smooth muscle cells, and interstitial cells of Cajal (ICCs), the ICCs have been thought to act as a pacemaker, and therefore it is important to decipher the cellular functions of ICCs to further our understanding of gut peristalsis. c‐Kit, a tyrosine kinase receptor, has widely been used as a marker for ICCs. Most studies with ICCs have been conducted in mammals using commercially available anti‐c‐Kit antibody. Recently, the chicken embryonic gut has emerged as a powerful model to study gut peristalsis. However, since the anti‐c‐Kit antibody for mammals does not work for chickens, cellular mechanisms by which ICCs are regulated have largely been unexplored. Here, we report a newly raised polyclonal antibody against the chicken c‐Kit protein. The specificity of the antibody was validated by both western blotting analyses and immunocytochemistry. Co‐immunostaining with the new antibody and anti‐α smooth muscle actin (αSMA) antibody successfully visualized ICCs in the chicken developing hindgut in the circular muscle and longitudinal muscle layers. As previously shown in mice, common progenitors of ICCs and smooth muscle cells at early stages were double positive for αSMA and c‐Kit, and at later stages, differentiated ICCs and smooth muscle cells exhibited only c‐Kit and αSMA, respectively. A novel ICC population was also found that radially extended from the submucosal layer to the circular muscle layer. Furthermore, the new antibody delineated individual ICCs in a cleared hindgut. The antibody newly developed in this study will facilitate the study of peristaltic movement in chicken embryos.