New monoclonal antibody (AIC) identifies interstitial cells of Cajal in the musculature of the mouse gastrointestinal tract
New monoclonal antibody (AIC) identifies interstitial cells of Cajal in the musculature of the mouse gastrointestinal tract
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DOI：
10.1016/j.autneu.2004.05.004
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发表时间：
2004-06-30
影响因子：
2.7
通讯作者：
Fujimoto, T
中科院分区：
文献类型：
--
作者：
Torihashi, S;Yokoi, K;Fujimoto, T
Cajal间质细胞（Interstitial cells of Cajal，ICC）是自发性肌肉收缩的起搏细胞，也是介导从肠道神经元到胃肠道平滑肌细胞的神经传递的神经调质。它们表达c-Kit，并且c-Kit的抗体（特别是ACK 2）已经成为功能和形态学研究的有用工具。然而，ACK 2对用多聚甲醛固定的组织不起作用，并且并非所有ICC在人体中都表达c-Kit。因此，为了找到新的ICC标记物和/或新的抗醛固定的抗体，我们制备了一种新的识别ICC的单克隆抗体，然后研究其抗原的性质。分离的ICC用于免疫。免疫组化筛选与骨髓瘤SP2融合的杂交瘤。在连续切片上应用ACK 2和每种抗体，并建立产生染色ICC的抗ICC抗体（AIC）的克隆。在W/Wv小鼠的其他器官和GI肌肉中检查AIC免疫阳性细胞的分布。用斑点杂交法研究其生化特性。AIC识别ICC，但免疫阳性细胞在W/Wv小鼠及其他器官的分布与c-Kit不同。多聚甲醛的免疫反应性是稳定的，但被Triton X-100或SDS阻断。结论：新抗体AIC识别ICC，但其抗原不是c-Kit，证实了除了c-Kit外，还存在良好的ICC标志物。虽然抗原尚未被分离，AIC是适合的形态学研究，是有用的调查ICC的c-Kit突变体。(C)2004 Elsevier B. V.保留所有权利。
Interstitial cells of Cajal (ICC) are pacemaker cells for the spontaneous muscular contractions and neuromodulators that mediate neurotransmission from enteric neurons to smooth muscle cells in the gastrointestinal (GI) tract. They express c-Kit, and the antibody for c-Kit (especially ACK2) has been a useful tool for functional and morphological studies. ACK2, however, does not work on tissues fixed with paraformaldehyde, and not all ICC express c-Kit in human. Therefore, in order to find a new marker of ICC and/or new antibody resisting aldehyde fixation, we produced a new monoclonal antibody that identifies ICC and then investigated the properties of its antigen. Isolated ICC were used for immunization. Hybridomas fused with myeloma SP2 were screened by immunohistochemistry. ACK2 and each antibody were applied on serial sections, and the clone producing anti-ICC antibody (AIC) that stains ICC was established. The distribution of AIC immunopositive cells was examined in other organs and also GI muscles of W/Wv mice. The biochemical properties were studied using dot blot analysis. AIC recognized ICC; however, distribution of immunopositive cells in W/Wv mice and other organs was different from that of c-Kit. The immunoreactivity was stable for paraformaldehyde but was blocked by either Triton X-100 or SDS. In conclusion, new antibody AIC recognized ICC but the antigen was not c-Kit, which confirms the existence of good markers of ICC besides c-Kit. Although the antigen has not been isolated, AIC is suitable for morphological study and is useful for investigation of ICC in c-Kit mutants. (C) 2004 Elsevier B.V. All rights reserved.