Hasegawa H, Lian S C, Finkbeiner W E, Verkman A S
Department of Medicine, University of California, San Francisco 94143-0521.
Am J Physiol. 1994 Apr;266(4 Pt 1):C893-903. doi: 10.1152/ajpcell.1994.266.4.C893.
This study is an extension of in situ hybridization experiments showing expression of mRNA encoding CHIP28 in selected epithelial or endothelia in spleen, colon, lung, and eye (H. Hasegawa, R. Zhang, A. Dohrman, and A. S. Verkman. Am. J. Physiol. 264 (Cell Physiol. 33): C237-C245, 1993). Additional tissues from rat were screened by in situ hybridization, and tissues from rat and humans were stained with a polyclonal anti-CHIP28 antibody. Northern blot showed the 2.8-kilobase mRNA encoding CHIP28 in kidney, lung, and heart. In situ hybridization showed strong hybridization in epithelial cells in choroid plexus, iris, ciliary body, and lens and in epithelial and subepithelial layers of trachea. Except for colonic crypts, specific hybridization was not observed in the gastrointestinal tract, liver, thyroid gland, and muscle. Immunoblot of tissues from exsanguinated rats showed immunoreactive CHIP28 protein in kidney, lung, trachea, and heart. In fixed frozen rat and/or human tissues, the anti-CHIP28 antibody stained epithelial cells in kidney proximal tubule and thin limb of Henle, lung alveolus, bronchial mucosa and glands, choroid plexus, ciliary body, iris, lens surface, colonic crypt, sweat gland, pancreatic acini, gallbladder epithelium, and placental syncytial trophoblast cells. Endothelial cells were stained in many tissues. These studies indicate a wide and selective CHIP28 tissue distribution, suggesting an important role for CHIP28 in fluid transport. The absence of CHIP28 in many nonrenal membranes believed to be water permeable suggests the existence of non-CHIP28 water transporters.
本研究是原位杂交实验的扩展,该实验显示编码CHIP28的mRNA在脾脏、结肠、肺和眼睛的特定上皮或内皮细胞中表达(长谷川H、张R、多尔曼A和韦克曼AS。《美国生理学杂志》264(细胞生理学33):C237 - C245,1993年)。通过原位杂交筛选了来自大鼠的其他组织,并用多克隆抗CHIP28抗体对来自大鼠和人类的组织进行染色。Northern印迹显示在肾脏、肺和心脏中有编码CHIP28的2.8千碱基mRNA。原位杂交显示在脉络丛、虹膜、睫状体和晶状体的上皮细胞以及气管的上皮和上皮下层有强烈杂交信号。除结肠隐窝外,在胃肠道、肝脏、甲状腺和肌肉中未观察到特异性杂交信号。对放血大鼠的组织进行免疫印迹显示,在肾脏、肺、气管和心脏中有免疫反应性CHIP28蛋白。在固定的冷冻大鼠和/或人类组织中,抗CHIP28抗体对肾近端小管和髓袢细段、肺泡、支气管黏膜和腺体、脉络丛、睫状体、虹膜、晶状体表面、结肠隐窝、汗腺、胰腺腺泡、胆囊上皮和胎盘合体滋养层细胞中的上皮细胞进行染色。许多组织中的内皮细胞也被染色。这些研究表明CHIP28在组织中的分布广泛且具有选择性,提示CHIP28在液体转运中起重要作用。许多被认为具有水通透性的非肾膜中不存在CHIP28,这表明存在非CHIP28水转运蛋白。