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Rat parietal cells express CCK(2) receptor mRNA: gene expression analysis of single cells isolated by laser-assisted
Karin Tømmerås1, Ingunn Bakke, Arne K Sandvik
1Department of Intra-Abdominal Diseases, Faculty of Medicine, The Norwegian University of Science and Technology, N-7489 Trondheim, Norway. karin.tommeras@medisin.ntnu.no
Abstract:
Gastrin plays a crucial role in maintaining a normal cellular composition and function of the oxyntic mucosa. It has been debated for decades whether parietal cells possess cholecystokinin-2 (CCK(2)) receptors and interact directly with gastrin. We investigated whether parietal cells express CCK(2) receptor mRNA by using new molecular biology techniques. Rat oxyntic mucosal cells were dispersed and enriched by elutriation, and single parietal and ECL cells were isolated from cell smears by means of laser microbeam microdissection and laser pressure catapulting. The mRNA from each single cell was isolated and subjected to one-step multiplex or conventional reverse transcription-polymerase chain reaction and subsequent nested PCR. Specific primers for the CCK(2) receptor were used in combination with primers for H,K-ATPase and histidine decarboxylase, specific markers for parietal and ECL cells, respectively. CCK(2) receptor mRNA was detected in 25% of the rat parietal cells and 40% of the ECL cells examined.
