Gastrin in the Perinatal Rat Pancreas and Gastric Antrum: Immunofluorescence Localization of Pancreatic Gastrin Cells and Gastrin Secretion in Monolayer Cell Cultures
The presence and development of immunoreactive gastrin (IRGa) in the fetal and neonatal pancreas and pyloric antrum of the rat were studied. IRGa appeared in both organs at least as early as the 16th day of fetal life. Antral IRGa increased rapidly and continuously in the neonatal period, while pancreatic IRGa concentration increased and was maintained at a relatively constant level from days 5 to 35. Monolayer cell cultures of the neonatal rat pancreas were used to evaluate the role of cyclic[c]AMP mediated release of gastrin. The addition of N6,O2''-dibutyryl cAMP (4 mM) or theophylline (4 mM) to the culture medium induced significant release of gastrin. The stimulation of adenylate cyclase with cholera toxin (10 ng/ml) also resulted in significant gastrin release. Long-term cultures (18-24 days) released gastrin continuously at a relative constant rate. The cellular localization of pancreatic gastrin in 7 day old cultures was performed by immunological techniques, using fluorescein-labeled antibodies to gastrin. The gastrin-containing cells were located at the periphery of most of the endocrine cell clusters. Immunofluorescence techniques for insulin and glucagon also showed that the .alpha. cells had a similar peripheral distribution, although they were more frequent in number. Insulin-containing cells were numerous and were present in all areas of the endocrine cell clusters. The studies support the following conclusions: gastrin is present in the rat pancreas, even as early as late fetal life; gastrin-producing cells are present and functionally competent in monolayer cell cultures of the neonatal rat pancreas for prolonged periods of time (24 days); gastrin is released from these cells when intracellular levels of cAMP are increased; by immunofluorescence methods, the gastrin-producing cells in pancreatic cell cultures are located at the periphery of the endocrine cell clusters.