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The Secretion of α40‐MSH from Xenopus Melanotropes Involves Calcium Influx through ω‐Conotoxin‐Sensitive Voltage‐Operated Calcium Channels
Author(s) -
Scheenen Wim J. J. M.,
Koning Harry P.,
Jenks Bruce G.,
Vaudry Hubert,
Roubos Eric W.
Publication year - 1994
Publication title -
journal of neuroendocrinology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.062
H-Index - 116
eISSN - 1365-2826
pISSN - 0953-8194
DOI - 10.1111/j.1365-2826.1994.tb00607.x
Subject(s) - calcium , endocrinology , voltage dependent calcium channel , medicine , t type calcium channel , depolarization , chemistry , voltage clamp , nifedipine , secretion , xenopus , biophysics , n type calcium channel , biology , membrane potential , biochemistry , gene
Abstract The secretory activity of endocrine cells largely depends on the concentration of free cytosolic calcium. We have studied the mechanisms that are involved in supplying the calcium necessary for the secretion of α‐melanophore‐stimulating hormone (α‐MSH) from melanotrope cells in the pituitary intermediate lobe of the amphibian Xenopus laevis . Using whole‐cell voltage clamp, high‐voltage activated calcium currents were observed, with a peak current between 0 and +20 mV. Two types of Ca 2 + ‐currents appeared, depending on the experimental setup. An inactivating current, which was observed after a 10 msec depolarizing prepulse, resembled currents through N‐type channels as it was clearly inhibited by 1 μM ω‐conotoxin. The second type was a non‐inactivating current, which was blocked up to 50% by 1 μM nifedipine, indicating its L‐type nature. Only a small component of this inactivating current could be blocked by ω‐conotoxin. No evidence was found for the presence of transient, low‐voltage activated currents. The spontaneous secretion of α‐MSH from superfused neurointermediate lobes was dependent on extracellular calcium, as low calcium conditions (10 −4 ‐10 −8 M) rapidly inhibited this process. Under these conditions, secretion was not affected by depolarizing concentrations of potassium chloride. The calcium ionophore A23187 increased secretion under low calcium conditions, but had no effect on spontaneous α‐MSH release. Treatment with CoCI 2 , a blocker of calcium channels, strongly inhibited the secretory process. These results suggest that spontaneous α‐MSH release depends on influx of calcium through voltage‐operated calcium channels. Nifedipine did not affect spontaneous secretion from lobes, nor did it affect potassium‐induced α‐MSH secretion from dispersed melanotropes. Also BAY‐K8644, a specific agonist of L‐type channels, did not influence α‐MSH release, neither under normal nor under low calcium conditions. On the other hand, ω‐conotoxin dose‐dependently inhibited α‐MSH release, to a maximum of 65% at a concentration of 5 μM, and inhibited potassiuminduced secretion by 40%. Thapsigargin, an agent that mobilizes calcium ions from intracellular stores, had no effect on spontaneous α‐MSH release under normal or low calcium conditions. From these results it is concluded that the spontaneous release of α‐MSH by melanotropes of X. laevis is effectuated by calcium influx through ω‐conotoxin‐sensitive, voltage‐operated N‐type calcium channels and that mobilization of calcium from intracellular stores does not play a major role in the regulation of this release.