cosyntropin and 2--5--dideoxyadenosine

ACTH (1-24) induces cell shape changes in the immunocytes of the bivalve mollusc, Mytilus galloprovincialis. Using computer-assisted microscopic image analysis, we have found that the G protein antagonist suramin sodium, the adenylate cyclase inhibitor 2',5'-dideoxyadenosine, and the protein kinase inhibitor staurosporine inhibit this effect. The highly specific inhibitors H-89 (for protein kinase A) and calphostin C (for protein kinase C) only inhibited partially the morphological alterations. In contrast, the simultaneous action of H-89 and calphostin C completely blocked these changes. The above findings indicate that ACTH (1-24) induces cell shape changes in molluscan immunocytes via adenylate cyclase/cAMP/protein kinase A pathway, as well as the activation of protein kinase C.

Topics: Adenylyl Cyclase Inhibitors; Animals; Bivalvia; Cell Size; Cosyntropin; Dideoxyadenosine; Enzyme Inhibitors; GTP-Binding Proteins; Hemocytes; Hemolymph; Image Processing, Computer-Assisted; Isoquinolines; Microscopy, Phase-Contrast; Naphthalenes; Protein Kinase Inhibitors; Signal Transduction; Staurosporine; Sulfonamides; Suramin

The steady-state relationship between the activation state of cAMP-dependent protein kinase (A-kinase) and lipolysis has been defined quantitatively. A-kinase activation was assessed by measuring the ( +/- cAMP) activity ratio in adipocyte extracts, and lipolysis was determined by measuring glycerol release from cells. Both processes were stimulated either by incubating cells in a ligand-free environment achieved with adenosine deaminase or by addition of lipolytic hormones. A response spectrum was obtained with a variety of adenylate cyclase stimulators and inhibitors, both receptor- and nonreceptor-mediated. Regardless of the ligands used to manipulate adipocyte activity, lipolysis varied from nil to maximal as the A-kinase activity ratio varied from approximately 0.05 to 0.3-0.35. These data provide a quantitative description of the steady-state relationship between A-kinase activity and lipolysis and indicate that the various lipolytic and antilipolytic agents tested act on the lipolytic process exclusively by altering adenylate cyclase activity and, thus, cellular cAMP concentrations. The data reveal also that transient "peaking" of cAMP, as measured by A-kinase activity ratios, is not an inherent feature of adipocyte metabolism. Moreover, the concentration requirements for lipolytic hormone action are critically dependent on the ambient concentration of antilipolytic agents, and t concentration requirements for antilipolytic agents are dependent on the extent to which cells are stimulated. The data in this paper provide the basis for assessing the relationship between A-kinase activity ratio and lipolysis in the presence of insulin (Londos, C., Honnor, R. C., and Dhillon, G. S. (1985) J. Biol. Chem. 260, 15139-15145).

Topics: 4-(3-Butoxy-4-methoxybenzyl)-2-imidazolidinone; Adipose Tissue; Animals; Colforsin; Cosyntropin; Deoxyadenosines; Dideoxyadenosine; Enzyme Activation; Glucagon; Glycerol; Isoproterenol; Kinetics; Lipolysis; Male; Phenylisopropyladenosine; Protein Kinases; Rats; Rats, Inbred Strains; Time Factors