| Objective: 1. To select a better fixative for immunohistochemically detecting histamine positive cells under light microscope. 2. To study the ontogeny, distribution, morphology and quantitative change of histamine positive cells in the mouse stomach from day 1 to day 45 after birth, as well as the anatomical relationship histamine positive cells with G cells, D cells, and parietal cells.Methods: the samples were fixed with 4%formaldehyde, Bouin's fluid, 4% paraformaldehyde and MAW fluid respectively in order to select a suitable fixative for the histamine immunochemical techniques and to define the most appropriate dilution of histamine antibody. The sections of mouse stomach from day 1 to day 45 after birth were stained with immunohistochemical SP method to show the ontogeny, distribution and morphological characteristics of hitamine positive cells in the stomach . The anatomical relationship of histamine positive cells with G cells, D cells and parietal cells was also studied using SP method.Results: 1. The sections of mouse stomach fixed with 4% formaldehyde or MAW fluid couldn't show histamine positive cells clearly, while fixed with Bouin's fluid, positive histamine granules were concentrated in the cytoplasm in a dissolved state. However, after fixed with 4% paraformaldehyde, histamine positive cells were shown clearly and the immunoreactive positive granules were concentrated in the cytoplasm. The most appropriate dilution of histamine antibody is 1:400, among ten different dilutions from 1:100 to 1:1000.2. Histamine positive cells could only be found in the mouse gastric mucosa as early as at day 5 after birth. At day 15 after birth, the number of histamine positive cells increased, and the cells existed alone or assembly. At day 30, 45 after birth, with increase in the thickness of gastric mucosa gradually and full developing of oxyntic gland, the number of histamine positive cells significantly increased. These cells were predominantly located in the basal third of the oxyntic mucosa, in a shape of "band". In the middle of oxyntic mucosa, histamine positive cells were seen scattered between parietal cells. A few histamine positive cells were found in the submucosa of both the antrum and the body of the stomach. Under the light microscope, the hstamine positive cells in the epithelium of the gastric mucosa were the closed type, which were small and were various in shape. The immunoreactive positive processes of these cells could be seen extending between adjacent other cells. The immunoreactive positive granules were stained deeply and found not only in the cytoplasm, but also in the extending slender process. It was often seen that histamine positive cells surrounding parietal cells. At the same time, some immunoreactive positive substance could be found around these cells.3. G cells were only found in the mucosa of the antrum, D cells were distributed in the mucosa of the body of the stomach, but it's number was significantly lower than that of histamine positive cells. Both kinds of cells extended their processes between adjacent other cells.Conclusion: 1. 4% paraformaldehyde is a suitable fixative for the histamine immunochemical technique, and the most appropriate dilution of hastamine antibody is 1:400. 2. The ontogeny of histamine positive cells in the mouse stomach was later than that of G cells, D cells and EC cells. The quantitative change of histamine positive cells was relevant intently with the development of gastric mucosa and the oxyntic gland. According to the distribution , these histamine positive cells located in the basal third oxyntic nucosa were ECL cells. 3. The relationship of ECL cells, G cells, D cells andparietal cells suggested that G cells secreted gastrin, which acted on ECL cells by way of endocrine. It's likely that histamine secreted by ECL cells had a endocrine or paracrine effect on parietal cells. And D cells regulated these three kinds of cells by way of paracrine probably. They were all involved in regulating the secret...
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