Abstract
Publisher Summary Permeabilization by hypoosmotic shock treatment is a gentle method for the rapid labeling of large quantities of clonal endocrine GH 3 cells with [ 3 H] inositol, a substrate for the synthesis of Phosphatidylinositol (Ptdlns). Further work will be required to establish the mechanism for the basal and thyrotropin-releasing hormone (TRH)-stimulated increases in the labeling of Ptdlns, which could occur through enzyme-mediated base exchange as well as through Ptdlns-synthase activity. Both activities have been found in GH 3 cell membranes. The hypo-osmotic shock treatment (HOST) method may be used for the studies of other metabolites in the inositol lipid pathway. For example, preliminary studies in the laboratory have shown increased incorporation of label into CDP-diglyceride after HOST permeabilization in the presence of [ 3 H] CTP. The true strength of this method is that the treated cells grow normally in culture, making it possible to study the metabolism of a wide range of introduced substrates under a variety of growth conditions.
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