Expression of somatostatin receptor subtypes on guinea pig gastric and colonic smooth muscle cells.
In vivo and in vitro studies have demonstrated that somatostatin can influence motility and smooth muscle contractility of the stomach and colon. Recent studies have proposed that some of these effects may be mediated by somatostatin receptors (sst) directly on the smooth muscle cells. If this is correct, the sst receptor subtypes that are present are unknown. This study aimed to resolve these points. Because nucleotide sequences of guinea pig sst genes are unknown, we used sst subtype-specific primers based on comparisons of human and rat sst subtypes and performed RT-PCR of DNase I-treated total RNA from guinea pig total brain. PCR products were cloned in pCR II and sequenced and showed 87% (sst(1)), 90% (sst(2)), 90% (sst(3)), 99% (sst(4)), and 80% (sst(5)), respectively, nucleotide homology to the same region (transmembrane 4-6) of the human sst genes. Homology to rat sequences were lower. PCR products were obtained from first-strand cDNA derived from DNase I-treated RNA from dispersed guinea pig gastric and colonic smooth muscle cells. In gastric and colonic smooth muscle cells, we detected sst(1)-sst(3) and sst(5), and all were confirmed by sequencing. The presence of sst(4) was shown by Southern blot analysis and hybridization with a guinea pig sst(4)-specific primer. RT-PCR from cultured colonic and gastric smooth muscle cells devoid of any neural elements gave identical results. These results demonstrate that in the guinea pig all five sst subtypes are present directly on gastric and colonic smooth muscle cells. Previous studies have suggested that a predominant sst(3) subtype on gastric and a sst(5) subtype on colonic muscle cells mediated somatostatin's contractile effects, but the finding here that all five sst subtypes exist on both of these cells suggests that other sst subtypes have only a small or no contractile effect, sst subtypes in guinea pig have a different pharmacological profile from rat or human sst, or these other sst subtypes have some yet undescribed physiological function in muscle cells.
Corleto
V D
VD
Digestive Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892-1804, USA.
Weber
H C
HC
Jensen
R T
RT
D000595:Amino Acid Sequence / Q000235:genetics; D000818:Animals; D015139:Blotting, Southern; D002478:Cells, Cultured; D003001:Cloning, Molecular; D003106:Colon / Q000166:cytology / Q000378:metabolism*; D005753:Gastric Mucosa / Q000378:metabolism*; D006168:Guinea Pigs; D007150:Immunohistochemistry; D008297:Male; D008969:Molecular Sequence Data; D009130:Muscle, Smooth / Q000166:cytology / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000378:metabolism; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D017481:Receptors, Somatostatin / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013270:Stomach / Q000166:cytology
D009419:Nerve Tissue Proteins; D020033:Protein Isoforms; D017481:Receptors, Somatostatin