Immunofluorescence detection of gastric H+/K+-ATPase and its alterations as related to acid secretion

Parietal cells in the rat oxyntic mucosa were analyzed by the immunofluorescence pattern of the proton pump. The adult rats were grouped into fasting (C), gastrin-treated (G), and ranitidine-treated (R) groups, gastric pH was measured, and the stomach was processed for immunohistochemistry. The fluorescence of parietal cells showed a reticular, diffuse, or mixed pattern in cytoplasm. Quantitatively, 53% of the total cells showed the reticular pattern in group G (pH 1.9), 44% in group C (pH 2.0), and 0% in group R (pH 6.7). On the other hand, 7.0%, of the total cells showed the diffuse pattern in group G, 11.9% in group C, and 56.2% in group R. The results indicated that the staining pattern depended on the activity of acid secretion. In addition, the proportion of parietal cells showing the reticular pattern decreased in the following order, the superficial, middle, and deep third of the mucosa, and the diffuse pattern showed the opposite trend. This suggests that the acid secretion is more active in parietal cells in the superficial part of the mucosa. The double staining with proton pump-specific and cytochrome oxidase-specific antibodies revealed the close relation between reticular fluorescence and mitochondria.