A current activated on depletion of intracellular Ca2+ stores can regulate exocytosis in adrenal chromaffin cells.
Exocytosis in excitable cells is strongly coupled to Ca2+ entry through voltage-gated channels but can be evoked by activation of membrane receptors that release Ca2+ from inositol 1,4, 5-trisphosphate-sensitive internal stores. In many cell types, depletion of Ca2+ stores activates Ca2+ influx across the plasma membrane, a process known as capacitative or store-operated Ca2+ entry. This influx is mediated by a number of voltage-independent, Ca2+-selective currents. In addition to replenishing Ca2+ stores, these currents are hypothesized to play an important role in agonist-evoked secretion in nonexcitable cells, although this has not been confirmed experimentally. The existence and physiological function of such currents in excitable cells is not known. Using the capacitance detection technique to monitor exocytosis, we provide direct experimental evidence that a similar mechanism exists in bovine adrenal chromaffin cells. Depletion of intracellular Ca2+ stores with thapsigargin, a SERCA pump inhibitor, or with BAPTA, an exogenous Ca2+ chelator, activates a small-amplitude, voltage-independent current that is carried by Ca2+ and Na+ ions. Ca2+ entry through this pathway is sufficient to stimulate exocytosis at negative membrane potentials. In addition, depolarization-evoked exocytosis is markedly facilitated on activation of the current. These data suggest that excitable cells possess a store-operated Ca2+ influx mechanism that may both directly trigger exocytosis and modulate excitation-secretion coupling.
Fomina
A F
AF
Department of Neurobiology and Anatomy, Medical College of Pennsylvania Hahnemann University, Philadelphia, Pennsylvania 19129, USA.
Nowycky
M C
MC
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002395:Catecholamines / Q000032:analysis; D002417:Cattle; D002614:Chelating Agents / Q000494:pharmacology; D019439:Chromaffin Cells / Q000187:drug effects / Q000378:metabolism*; D004533:Egtazic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D004553:Electric Conductivity; D005089:Exocytosis / Q000187:drug effects / Q000502:physiology*; D008081:Liposomes; D008564:Membrane Potentials / Q000187:drug effects; D018408:Patch-Clamp Techniques; D019284:Thapsigargin / Q000494:pharmacology
D002395:Catecholamines; D002614:Chelating Agents; D008081:Liposomes; D004533:Egtazic Acid; D019284:Thapsigargin; C025603:1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid; D002118:Calcium