Evidence that the ability to respond to a calcium stimulus in exocytosis is determined by the secretory granule membrane: comparison of exocytosis of injected bovine chromaffin granule membranes and endogenous cortical granules in Xenopus laevis oocytes.
1. To understand better the mechanisms which govern the sensitivity of secretory vesicles to a calcium stimulus, we compared the abilities of injected chromaffin granule membranes and of endogenous cortical granules to undergo exocytosis in Xenopus laevis oocytes and eggs in response to cytosolic Ca2+. Exocytosis of chromaffin granule membranes was detected by the appearance of dopamine-beta-hydroxylase of the chromaffin granule membrane in the oocyte or egg plasma membrane. Cortical granule exocytosis was detected by release of cortical granule lectin, a soluble constituent of cortical granules, from individual cells. 2. Injected chromaffin granule membranes undergo exocytosis equally well in frog oocytes and eggs in response to a rise in cytosolic Ca2+ induced by incubation with ionomycin. 3. Elevated Ca2+ triggered cortical granule exocytosis in eggs but not in oocytes. 4. Injected chromaffin granule membranes do not contribute factors to the oocyte that allow calcium-dependent exocytosis of the endogenous cortical granules. 5. Protein kinase C activation by phorbol esters stimulates cortical granule exocytosis in both Xenopus laevis oocytes and X. laevis eggs (Bement, W. M., and Capco, D. G., J. Cell Biol. 108, 885-892, 1989). Activation of protein kinase C by phorbol ester also stimulated chromaffin granule membrane exocytosis in oocytes, indicating that although cortical granules and chromaffin granule membranes differ in calcium responsiveness, PKC activation is an effective secretory stimulus for both. 6. These results suggest that structural or biochemical characteristics of the chromaffin granule membrane result in its ability to respond to a Ca2+ stimulus. In the oocytes, cortical granule components necessary for Ca(2+)-dependent exocytosis may be missing, nonfunctional, or unable to couple to the Ca2+ stimulus and downstream events.
Scheuner
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Department of Pharmacology, University of Michigan Medical School, Ann Arbor 48109-0632.
Holz
R W
RW
D000818:Animals; D002118:Calcium / Q000494:pharmacology*; D002837:Chromaffin Granules / Q000378:metabolism*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D003594:Cytoplasmic Granules / Q000378:metabolism*; D004299:Dopamine beta-Hydroxylase / Q000378:metabolism; D004533:Egtazic Acid / Q000494:pharmacology; D005089:Exocytosis / Q000187:drug effects* / Q000502:physiology; D005260:Female; D007425:Intracellular Membranes / Q000187:drug effects / Q000378:metabolism; D015759:Ionomycin / Q000494:pharmacology; D037102:Lectins / Q000378:metabolism; D009865:Oocytes / Q000166:cytology / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014982:Xenopus laevis
D037102:Lectins; D004533:Egtazic Acid; D015759:Ionomycin; D004299:Dopamine beta-Hydroxylase; D017868:Cyclic AMP-Dependent Protein Kinases; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium