Gomez-Pinilla Pedro J, Gibbons Simon J, Bardsley Michael R, Lorincz Andrea, Pozo Maria J, Pasricha Pankaj J, Van de Rijn Matt, West Robert B, Sarr Michael G, Kendrick Michael L, Cima Robert R, Dozois Eric J, Larson David W, Ordog Tamas, Farrugia Gianrico
Enteric Neuroscience Program, Miles and Shirley Fiterman Center for Digestive Diseases and Department of Physiology and Biomedical Engineering, Mayo Clinic, 200 First St. SW, Rochester, MN 55905, USA.
Am J Physiol Gastrointest Liver Physiol. 2009 Jun;296(6):G1370-81. doi: 10.1152/ajpgi.00074.2009. Epub 2009 Apr 16.
Populations of interstitial cells of Cajal (ICC) are altered in several gastrointestinal neuromuscular disorders. ICC are identified typically by ultrastructure and expression of Kit (CD117), a protein that is also expressed on mast cells. No other molecular marker currently exists to independently identify ICC. The expression of ANO1 (DOG1, TMEM16A), a Ca(2+)-activated Cl(-) channel, in gastrointestinal stromal tumors suggests it may be useful as an ICC marker. The aims of this study were therefore to determine the distribution of Ano1 immunoreactivity compared with Kit and to establish whether Ano1 is a reliable marker for human and mouse ICC. Expression of Ano1 in human and mouse stomach, small intestine, and colon was investigated by immunofluorescence labeling using antibodies to Ano1 alone and in combination with antibodies to Kit. Colocalization of immunoreactivity was demonstrated by epifluorescence and confocal microscopy. In the muscularis propria, Ano1 immunoreactivity was restricted to cells with the morphology and distribution of ICC. All Ano1-positive cells in the muscularis propria were also Kit positive. Kit-expressing mast cells were not Ano1 positive. Some non-ICC in the mucosa and submucosa of human tissues were Ano1 positive but Kit negative. A few (3.2%) Ano1-positive cells in the human gastric muscularis propria were labeled weakly for Kit. Ano1 labels all classes of ICC and represents a highly specific marker for studying the distribution of ICC in mouse and human tissues with an advantage over Kit since it does not label mast cells.
在几种胃肠道神经肌肉疾病中, Cajal间质细胞(ICC)群体发生了改变。ICC通常通过超微结构和Kit(CD117)的表达来鉴定,Kit是一种也在肥大细胞上表达的蛋白质。目前不存在其他分子标记物来独立鉴定ICC。胃肠道间质瘤中钙激活氯离子通道ANO1(DOG1,TMEM16A)的表达表明它可能作为ICC标记物有用。因此,本研究的目的是确定与Kit相比Ano1免疫反应性的分布,并确定Ano1是否是人和小鼠ICC的可靠标记物。使用单独的抗Ano1抗体以及与抗Kit抗体联合,通过免疫荧光标记研究Ano1在人和小鼠胃、小肠和结肠中的表达。通过落射荧光显微镜和共聚焦显微镜证实免疫反应性的共定位。在固有肌层中,Ano1免疫反应性仅限于具有ICC形态和分布的细胞。固有肌层中所有Ano1阳性细胞也为Kit阳性。表达Kit的肥大细胞不是Ano1阳性。人组织黏膜和黏膜下层中的一些非ICC是Ano1阳性但Kit阴性。人胃固有肌层中少数(3.2%)Ano1阳性细胞对Kit标记较弱。Ano1标记所有类型的ICC,是研究小鼠和人组织中ICC分布的高度特异性标记物,相对于Kit具有优势,因为它不标记肥大细胞。