Effects of Insulin on the Synthesis, Intracellular Degradation, and Secretion of Parathormone*

Abstract

Bovine parathyroid organoids were maintained for up to 12 days of culture in the presence or absence of insulin. Insulin-treated organoids secreted more PTH and secretory protein-I (SP-I) than did untreated organoids at both 1.4 and 1.8 mM Ca, concentrations chosen to promote partially elevated and suppressed secretion rates, respectively. The insulin effect was dose dependent and reversible. To determine whether insulin might increase secretion by reducing degradation of cellular PTH, its effects on several parameters related to degradative processes were examined. Compared to control cultures maintained at either 1.4 or 1.8 mM Ca, insulin did not induce changes in the relative levels of intact hormone and COOH-terminal peptide fragments secreted into culture medium, nor did it decrease the total cellular levels of three lysosomal enzymes or mute the effects of 3-methyladenine (an agent that decreases formation of autophagosomes) to increase PTH secretion. Thus, insulin did not appear to increase PTH secretion by reducing the latter's cellular degradation. Synthesis of total proteins and of the secreted proteins SP-I and PTH was examined using short incubations of control and insulin-treated organoids with [3H] leucine. Incorporation of 3H into total acid-precipitable proteins was not elevated in insulin-treated organoids; that into PTH/pro-PTH and SP-I, however, was significantly greater in insulin-treated than in control organoids. The results suggested that the insulin-mediated increase in PTH and SP-I secretion is largely due to its regulation of PTH and SP-I biosynthesis.