guanylyl-imidodiphosphate and xanthosine-5--triphosphate

The role of GTP-binding proteins in exocytosis in bovine adrenal chromaffin cells was examined using patch-clamp capacitance measurement. Internal dialysis with the non-hydrolysable GTP analogue guanosine 5'-[beta gamma-imido]triphosphate and xanthosine triphosphate (XTP) activated a capacitance increase. Exocytosis triggered by XTP was blocked by guanosine 5'-[beta-thio]diphosphate (GDP beta S) but Ca(2+)-induced exocytosis was unaffected. The capacitance increase due to XTP could not be explained by Ca2+ mobilisation since Ins(1,4,5)P3 and caffeine did not mimic the response. Chromaffin cells appear to possess a Ca(2+)-independent pathway for exocytosis that involves GTP-binding proteins. The magnitude of the response to XTP suggested that GTP analogues stimulate both exocytosis and recruitment of secretory granules.

Topics: Adenosine Triphosphate; Adrenal Glands; Animals; Calcium; Cattle; Chromaffin System; Electric Conductivity; Exocytosis; GTP-Binding Proteins; Guanosine 5'-O-(3-Thiotriphosphate); Guanosine Diphosphate; Guanosine Triphosphate; Guanylyl Imidodiphosphate; Ribonucleotides; Thionucleotides

The effect of GTP analogues on catecholamine secretion and [3H]arachidonic acid release from digitonin-permeabilized adrenal chromaffin cells was examined. Several GTP analogues stimulated Ca2(+)-independent exocytosis, with the order of efficacy being XTP greater than ITP greater than guanosine 5'-[beta gamma-imido]triphosphate (p[NH]ppG) greater than guanosine 5'-[gamma-thio]triphosphate (GTP[S]). The stimulatory effect of the GTP analogues appeared to be due to activation of a conventional GTP-binding protein, as it was inhibited by guanosine 5'-[beta-thio]diphosphate (GDP[S]). In contrast, Ca2(+)-dependent exocytosis was only partially inhibited by high doses of GDP[S]. GTP did not stimulate Ca2(+)-independent exocytosis, but instead was found to inhibit secretion caused by micromolar Ca2+. Arachidonic acid (100 microM) also stimulated Ca2(+)-independent catecholamine secretion. Determination of the effect of GTP analogues on release of free [3H]arachidonic acid into the medium showed that it was stimulated by GTP[S] but inhibited by GTP, p[NH]ppG, ITP and XTP. The inhibition of [3H]arachidonic acid release by XTP was not prevented by GDP[S]. These results demonstrate that activation of a GTP-binding protein by certain GTP analogues can induce Ca2(+)-independent secretion in adrenal chromaffin cells and that the effect of GTP analogues on Ca2(+)-independent secretion can be dissociated from generation of arachidonic acid.

Topics: Adrenal Medulla; Animals; Arachidonic Acid; Arachidonic Acids; Calcium; Catecholamines; Cattle; Cell Membrane Permeability; Chromaffin System; Digitonin; Exocytosis; Guanosine 5'-O-(3-Thiotriphosphate); Guanosine Triphosphate; Guanylyl Imidodiphosphate; Inosine Triphosphate; Ribonucleotides; Thionucleotides