Expression of somatostatin receptor genes and their role in inhibiting Cl- secretion in HT-29cl.19A colonocytes.
Recent studies in a cultured model of the intestinal epithelium (HT-29cl.19A) have shown that somatostatin-14 (SS-14) inhibits the Cl- secretory process by acting at multiple G protein-dependent sites. These actions may underlie the antidiarrheal properties of SS peptides. This study has investigated the expression of specific SS receptor subtypes (SSTR) in HT-29cl.19A and examined their role in mediating SS antisecretory actions. Two predominant SSTR, SSTR1 and SSTR2, were identified by reverse transcriptase-polymerase chain reaction (RT-PCR) of mRNA from polarized HT-29cl.19A monolayers. Receptor binding studies showed evidence of two distinct populations of binding sites consistent with the known properties of SSTR1 and SSTR2. The role of SSTR in inhibition of secretion was investigated by comparing the effectiveness of native and synthetic SS peptides on adenosine 3',5'-cyclic monophosphate (cAMP)-dependent Cl- secretion. Secretion stimulated by the receptor-mediated agonist prostaglandin E2 (PGE2) was inhibited > 70% by SS-14 with a 50% effective concentration (EC50) of 32 nM. In contrast, SMS-201-995 (SMS) and RC-160 exhibited little or no antisecretory activity (maximum inhibition of 15 +/- 1.9 and 2.8 +/- 1.9%, respectively, at 100 microM; EC50 > 1.5 microM). Similar effects on PGE2-stimulated cAMP accumulation were also observed. SS-14, but not SMS, also inhibited secretion stimulated by dibutyryl cAMP, which acts independently of changes in cellular cAMP. Pretreatment with pertussis toxin reversed the antisecretory effects of SS peptides.(ABSTRACT TRUNCATED AT 250 WORDS)
D001483:Base Sequence; D002460:Cell Line; D002712:Chlorides / Q000037:antagonists & inhibitors* / Q000378:metabolism; D003106:Colon / Q000166:cytology / Q000378:metabolism* / Q000502:physiology*; D000242:Cyclic AMP / Q000378:metabolism / Q000494:pharmacology; D015870:Gene Expression; D007477:Ions; D015335:Molecular Probes / Q000235:genetics; D008969:Molecular Sequence Data; D017481:Receptors, Somatostatin / Q000235:genetics* / Q000502:physiology; D013004:Somatostatin / Q000031:analogs & derivatives / Q000138:chemical synthesis
D002712:Chlorides; D007477:Ions; D015335:Molecular Probes; D017481:Receptors, Somatostatin; D013004:Somatostatin; D000242:Cyclic AMP