Evidence for biosynthesis of preproinsulin in gut of rat.
Glucagon and other pancreatic peptides are made in the gut, but there is little evidence for the formation of insulin. The demonstration of insulin receptors on the mucosa of gut epithelium suggests that there may be an autocrine or paracrine role for insulin made in the gut. Such insulin may control cell division, the secretion of other peptides from the same or neighboring cells, or motility and absorption. To search for the ability of the gut to make insulin, sections of freshly excised segments of rat gut were treated with an antiserum against porcine insulin. Intracellular immunoreactivity appeared in glandular cells in the stomach and colon but not in the small intestine. Preproinsulin mRNA was detected in similar cells in the stomach and colon by in situ hybridization, using specific oligonucleotide probes. Rat preproinsulin 1 and 2 mRNAs were transcribed by reverse transcriptase to the corresponding cDNAs, which were then amplified by polymerase chain reaction, utilizing specific oligonucleotide primers. Restriction analysis confirmed the identity of rat preproinsulin 1 and 2 mRNA in the colon and rat preproinsulin 1 mRNA in the stomach. Neither was found in the small intestine. Base sequences of the cDNAs were identical to the coding regions of pancreatic rat preproinsulin 1 and 2 messages. These observations are strong evidence for the synthesis of preproinsulin in the gut of the rat.
Kendzierski
K S
KS
Department of Anatomy and Neurobiology, Medical College of Ohio, Toledo, 43614, USA.
Pansky
B
B
Budd
G C
GC
Saffran
M
M
D000818:Animals; D001483:Base Sequence; D003106:Colon / Q000737:chemistry; D004262:DNA Restriction Enzymes; D018076:DNA, Complementary / Q000737:chemistry; D004064:Digestive System / Q000737:chemistry / Q000378:metabolism*; D005260:Female; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D007328:Insulin; D007421:Intestine, Small / Q000737:chemistry; D008297:Male; D015345:Oligonucleotide Probes; D010179:Pancreas / Q000737:chemistry; D011384:Proinsulin / Q000096:biosynthesis* / Q000235:genetics; D011498:Protein Precursors / Q000096:biosynthesis* / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013270:Stomach / Q000737:chemistry
D018076:DNA, Complementary; D007328:Insulin; D015345:Oligonucleotide Probes; D011498:Protein Precursors; D012333:RNA, Messenger; C013737:preproinsulin; D011384:Proinsulin; D004262:DNA Restriction Enzymes