Tritiated Thymidine Incorporation into DNA in Rat Gastric Mucosal Cells

Abstract

The parietal cell acid pump inhibitor, omeprazole, has undergone numerous genotoxicity studies, the conclusions of which have all been negative. A recent report by Burlinson described a method which is claimed to measure unscheduled DNA synthesis (UDS) in gastric mucosa, based on the incorporation of tritiated thymidine (3H-TdR) into DNA. In a subsequent letter it was concluded that omeprazole induces a significant increase in UDS, and it was suggested that 'for omeprazole, a genotoxic action cannot be discounted'. The Burlinson method of measuring UDS relies upon selective separation of non-dividing oxyntic mucosal surface cells from the stem cells located in the proliferative zone. These stem cells undergo normal cell division, subsequently migrate to the top and bottom of the gland, differentiating into the various specialized cells of the gland such as surface, parietal and chief cells. In the oxyntic mucosa the majority of stem cells are located at a level corresponding to one fifth of the mucosal thickness from the surface. The purpose of this study was to evaluate the Burlinson method, which depends upon the selective isolation of non-dividing surface cells by pronase digestion. The fraction used for measurement was investigated in terms of 3H-TdR uptake, the cellular distribution of 3H-TdR by autoradiography, the number of parietal cells and the sensitivity of 3H-TdR uptake for hydroxyurea (an inhibitor of normal, scheduled DNA synthesis). The results show that pronase digestion did not selectively isolate surface epithelial cells and that the digest contained both parietal cells (15 +/- 1.5%) and glandular fragments.(ABSTRACT TRUNCATED AT 250 WORDS)