Haragsim L, Bastani B
Division of Nephrology, St. Louis University Health Sciences Center, Missouri 63110, USA.
Histochem J. 1996 Feb;28(2):117-22. doi: 10.1007/BF02331416.
The presence and distribution of gastric-type H(+)-K(+)-ATPase were investigated in the rat colon using a monoclonal antibody raised against hog gastric H(+)-K(+)-ATPase. Rat stomach was used as positive control. Rat kidney and ileum, in both of which H(+)-K(+)-ATPase has been reported in the past, were also studied. In stomach, very strong staining was found confined to the parietal cells, and a strong band at M(r) approximately 94 kDa on the immunoblots. In colon a moderate staining was found in the supranuclear region of the epithelial cells, with similar intensity and distribution of staining of the surface and deep mucosa of the crypts, throughout the length of the colon. Another monoclonal antibody, specific to the 31 kDa subunit of H(+)-ATPase, used as a negative control, or omission of the primary antibody, resulted in lack of any staining in either colon or stomach. On immunoblots of homogenates of colonic mucosa, no specific band could be identified, either due to very low expression of the H(+)-K(+)-ATPase or loss of antigenicity of the epitope during the processing steps. No positive staining was observed in rat kidney and ileum, suggesting that they contain isoforms that are structurally different.
使用针对猪胃H(+)-K(+)-ATP酶产生的单克隆抗体，研究大鼠结肠中胃型H(+)-K(+)-ATP酶的存在及分布情况。以大鼠胃作为阳性对照。还对大鼠肾脏和回肠进行了研究，过去已有报道称这二者中存在H(+)-K(+)-ATP酶。在胃中，发现极强的染色仅限于壁细胞，免疫印迹上在分子量约为94 kDa处有一条强条带。在结肠中，上皮细胞的核上区域有中度染色，在结肠全长的隐窝表面和深部黏膜中染色强度及分布相似。另一种针对H(+)-ATP酶31 kDa亚基的单克隆抗体用作阴性对照，或者省略一抗，结果结肠和胃均未出现任何染色。在结肠黏膜匀浆的免疫印迹上，由于H(+)-K(+)-ATP酶表达极低或在处理步骤中表位抗原性丧失，无法鉴定出特异性条带。在大鼠肾脏和回肠中未观察到阳性染色，表明它们含有的同工型在结构上有所不同。
