Combined non-radioactive detection of peptide hormones and their mRNAs in stomach somatostatin cells.
Non-radioactive in situ hybridization (ISH) and immunocytochemistry (ICC) have been used to detect somatostatin (SS) messenger RNA (mRNA) and peptide in antropyloric mucosa of the stomach in the rats. We have applied a method of non-radioactive in situ hybridization histochemistry using digoxigenin labelled oligonucleotide probes to detect somatostatin gene expression in the stomach. In prehybridization stage we used proteinase K (PK) in various concentrations (from 1 to 10 micrograms/ml) and periods (from 10 min to 1 h) but we maintained high background. However it was possible to detect the somatostatin mRNAs in the stomach mucosa making use of either background preventing solutions during the prehybridization, or of levamisole (20 microliters/mg) added into the hybridization buffer or of pepsin. Somatostatin mRNA and peptide signals were scattered all through the mucosa especially localized particularly at the base of the pyloric glands. SS peptide shown by ICC and SS mRNA shown by ISH were observed in different cells.
Bolkent
S
S
Department of Medical Biology, Cerrahpasa Faculty of Medicine, University of Istanbul, Istanbul 34303, Turkey.
Yilmazer
S
S
Ozturk
M
M
D000818:Animals; D005260:Female; D007150:Immunohistochemistry / Q000379:methods; D017403:In Situ Hybridization / Q000379:methods; D007413:Intestinal Mucosa / Q000737:chemistry / Q000166:cytology*; D036361:Peptide Hormones / Q000032:analysis* / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013004:Somatostatin / Q000032:analysis*
D036361:Peptide Hormones; D012333:RNA, Messenger; D013004:Somatostatin