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Volume 272, Number 13, Issue of March 28, 1997 pp. 8325-8331
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

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Effects of External Osmotic Pressure on Vesicular Secretion from Bovine Adrenal Medullary Cells

(Received for publication, November 11, 1996, and in revised form, January 13, 1997)

Ricardo Borges , Eric R. Travis ^§ , Spencer E. Hochstetler ^§ and R. Mark Wightman ^§

From the  Unidad de Farmacología, Facultad de Medicina, Universidad de La Laguna, 38010 Santa Cruz de Tenerife, Spain and the ^§ Department of Chemistry, University of North Carolina, Chapel Hill, North Carolina 27599-3290

Secretion of catecholamines from individual vesicles of bovine adrenal medullary cells was studied with amperometry in media of various osmolarities and compared with results obtained in isotonic physiological buffer (315 mosM). Hypotonic solutions caused an increase in the number of amperometric spikes evoked by brief exposure to 5 mM Ba²⁺. Under moderate hypertonic conditions (630 mosM), individual vesicular events were decreased in frequency, and lower amounts were secreted per event. Furthermore, the events were temporally broadened relative to those observed during release in isotonic conditions. At 970 mosM, exposure to 5 mM Ba²⁺ evoked even smaller secretory events that resemble the prespike feature that has been attributed to the initial opening of the fusion pore. The lack of large spikes is not due to failure of Ba²⁺ entry because fura-2 fluorescence reveals an increase in intracellular divalent ions. After exposure to Ba²⁺ in hypertonic solution, spikes could be induced with isotonic solution transiently directed onto the cell, but this process was not accompanied by a change in the concentration of intracellular divalent ions. Thus, this procedure provides an unique opportunity to temporally separate exocytotic secretion from entry of divalent ions.

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