Stimulation of adenylate cyclase from isolated hepatocytes and Kupffer cells.
Hepatocytes and Kupffer cells were separated from rat liver after prelabeling the Kupffer cells with colloidal iron and perfusion of the liver with digestive enzymes. The activity of several enzymes from Kupffer cells and hepatocytes was compared to validate this method of cell separation. The ratios of hepatocyte to Kupffer cell specific activities of glucose-6-phosphatase, 5'-nucleotidase, adenylate cyclase, and acid phosphatase were 20, 0.39, 0.18, and 0.078, respectively. Adenylate cyclases from hepatocytes and Kupffer cells were stimulated by fluoride ion, GTP, and catecholamines. Hepatocyte adenylate cyclase was also stimulated by glucagon, secretin, vasoactive intestinal polypeptide, and by prostaglandin E1, whereas, the Kupffer cell enzyme was completely insensitive to these hormones. The stimulation of hepatocyte adenylate cyclase by combinations of glucagon plus secretin, or glucagon plus vasoactive intestinal polypeptide, were equivalent to the sum of the individual stimulations. This suggests that the hepatocyte has specific receptors for glucagon and for vasoactive intestinal polypeptide and secretin. Prostaglandin E1 stimulation of hepatocyte adenylate cyclase was not additive to the stimulation caused by polypeptide hormones or catecholamines, nor did prostaglandin E1 decrease stimulation caused by these hormones. Although prostaglandin-sensitive adenylate cyclase was recovered with hepatocytes, 40 to 50% of the total liver prostaglandin-sensitive activity was recovered in a fraction of cell debris mixed with small cells which did not phagocytize colloidal iron.
D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D005459:Fluorides / Q000494:pharmacology; D005934:Glucagon / Q000494:pharmacology; D006160:Guanosine Triphosphate / Q000494:pharmacology; D006821:Hyaluronoglucosaminidase / Q000494:pharmacology; D066298:In Vitro Techniques; D007501:Iron / Q000494:pharmacology; D007545:Isoproterenol / Q000494:pharmacology; D007728:Kupffer Cells / Q000187:drug effects / Q000201:enzymology* / Q000648:ultrastructure; D008099:Liver / Q000187:drug effects / Q000201:enzymology* / Q000648:ultrastructure; D008297:Male; D003012:Microbial Collagenase / Q000494:pharmacology; D008858:Microscopy, Phase-Contrast; D010455:Peptides / Q000494:pharmacology; D010477:Perfusion; D011458:Prostaglandins E / Q000494:pharmacology; D051381:Rats; D012633:Secretin / Q000494:pharmacology
D010455:Peptides; D011458:Prostaglandins E; D012633:Secretin; D006160:Guanosine Triphosphate; D005934:Glucagon; D007501:Iron; D006821:Hyaluronoglucosaminidase; D003012:Microbial Collagenase; D000262:Adenylyl Cyclases; D007545:Isoproterenol; D005459:Fluorides; D002118:Calcium