Synthesis and secretion of somatostatin-28 and -14 by fetal rat intestinal cells in culture

Abstract

A fetal rat intestinal cell (FRIC) culture model was established to investigate the factors controlling synthesis and secretion of somatostatin-28 (S-28) by the intestine. Immunohistochemical analysis demonstrated the presence of cells containing somatostatin-like immunoreactivity (SLI), many of which emitted long processes toward neighboring cells. Gel chromatography of SLI stored and secreted by nonstimulated FRIC cultures demonstrated a predominance of S-28 (59%), lesser amounts of S-14 (31%), and a minor peak of large molecular weight SLI (10%). Secretion of SLI was stimulated by treatment of cells for 2 h with 5 mM dibutyryl adenosine 3',5'-cyclic monophosphate (DBcAMP; P less than 0.01) or 2 microM phorbol ester (P less than 0.05), but was unaffected by the calcium ionophore A23187 (2 micrograms/ml). Concomitant treatment with all three agents increased SLI secretion in an additive fashion to 254 +/- 25% of controls (P less than 0.0001). DBcAMP treatment did not alter the distribution of stored or secreted S-28 (59%) and S-14 (33%). After 24 h of exposure to DBcAMP, but not after treatment with phorbol ester, a threefold increment in prosomatostatin mRNA transcript levels was observed. FRIC cultures therefore synthesize and secrete a predominance of S-28 in a regulated manner, making them a promising model for future studies on the biosynthesis and secretion of intestinal somatostatin.