Gastrin effects on isolated rat enterochromaffin-like cells in primary culture.
The hormone gastrin stimulates acid secretion by releasing histamine from gastric enterochromaffin-like (ECL) cells and induces ECL cell proliferation in vivo. This study uses a > 90% pure ECL cell preparation in culture to compare gastrin effects on histamine release, histidine decarboxylase (HDC) activity, and DNA synthesis. Gastrin and the cholecystokinin octapeptide (CCK-8, nonsulfated) induced histamine release from ECL cells (24-96 h of primary culture) within 5 min of incubation [concentration eliciting 50% of maximal response (EC50), 4 and 2 x 10(-11) M, respectively]. The CCK-B antagonist L-365,260 inhibited this effect [concentration inhibiting 50% of maximal response (IC50), 2 x 10(-8) M], whereas the CCK-A antagonist L-364,718 (10(-8) M) and the tyrosine kinase inhibitor genistein (10(-4) M) had no effect. Histamine release was associated with a biphasic elevation of intracellular Ca2+. Gastrin stimulated HDC activity two- to threefold after 60 min of incubation (EC50, 10(-10) M). Gastrin also increased DNA synthesis in ECL cells, with an EC50 of 1.7 x 10(-12) M as measured by the incorporation of 5-bromo-2'-deoxyuridine (BrdU). Positive nuclear immunostaining increased two- to threefold in up to 20% of ECL cells after 48-96 h of incubation. This effect was inhibited by L-365,260 (IC50, 5 x 10(-9) M) and by genistein (10(-4) M) but was not altered by L-364,718 (10(-8) M). The antisecretory drugs omeprazole, lansoprazole, and pantoprazole did not affect BrdU incorporation in isolated ECL cells. In conclusion, acute and chronic gastrin effects on the ECL cell are mediated via CCK-B receptors but differ in apparent receptor affinity and signal transduction pathways.
Prinz
C
C
Department of Physiology, University of California, Los Angeles.
Scott
D R
DR
Hurwitz
D
D
Helander
H F
HF
Sachs
G
G
D000818:Animals; D001562:Benzimidazoles / Q000494:pharmacology; D001973:Bromodeoxyuridine / Q000493:pharmacokinetics; D002118:Calcium / Q000378:metabolism; D002469:Cell Separation; D002478:Cells, Cultured; D002766:Cholecystokinin / Q000037:antagonists & inhibitors; D004247:DNA / Q000096:biosynthesis; D004759:Enterochromaffin Cells / Q000187:drug effects* / Q000378:metabolism; D005260:Female; D005755:Gastrins / Q000494:pharmacology*; D006633:Histamine Antagonists / Q000494:pharmacology; D006636:Histamine Release; D006640:Histidine Decarboxylase / Q000378:metabolism; D007425:Intracellular Membranes / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley
D001562:Benzimidazoles; D005755:Gastrins; D006633:Histamine Antagonists; D004247:DNA; D002766:Cholecystokinin; D006640:Histidine Decarboxylase; D001973:Bromodeoxyuridine; D002118:Calcium