The readily releasable pool of vesicles in chromaffin cells is replenished in a temperature-dependent manner and transiently overfills at 37 degrees C.
Maturation of exocytic vesicles to the release-ready state is regulated by several factors, including intracellular calcium concentration ([Ca(2+)](int)) and the state of protein phosphorylation. Here we investigated the effects of temperature on the recovery from depletion of the readily releasable pool (RRP) of vesicles in adrenal chromaffin cells. Exocytosis and [Ca(2+)](int) were monitored by combined membrane capacitance and fura-2 measurements. At higher temperatures, a faster pool refilling and a larger RRP size were observed. The time constants of the recovery from depletion ranged from 3.6 to 1.1 sec (22 and 37 degrees C, respectively) yielding a Q(10) of 2.3. The changes of the Ca(2+) signal between the different temperatures could not account for the differences in recovery kinetics. At 32 and 37 degrees C, we observed a transient overfilling of the RRP after pool depletion, which stands in clear contrast to the sustained secretory depression seen at lower temperatures. The overshoot in RRP size was very prominent in cells with lower basal [Ca(2+)](int), hence with a large difference between prestimulus and poststimulus [Ca(2+)](int). In cells with higher basal [Ca(2+)](int), the pool was larger under steady-state conditions but showed less overfilling on stimulation. We conclude that vesicle maturation is markedly accelerated at physiological temperature, thus allowing for a rapid adaptation of the pool size to the relatively short-lived Ca(2+) transient.
Dinkelacker
V
V
Department of Membrane Biophysics, Max-Planck-Institute for Biophysical Chemistry, D-37077 Göttingen, Germany. vdinkel@gwdg.de
Voets
T
T
Neher
E
E
Moser
T
T
D000818:Animals; D002118:Calcium / Q000378:metabolism; D020013:Calcium Signaling / Q000502:physiology; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D019439:Chromaffin Cells / Q000166:cytology / Q000378:metabolism*; D004553:Electric Conductivity; D005089:Exocytosis / Q000502:physiology; D005456:Fluorescent Dyes; D016257:Fura-2 / Q000031:analogs & derivatives*; D007424:Intracellular Fluid / Q000378:metabolism; D018408:Patch-Clamp Techniques; D011930:Reaction Time / Q000502:physiology; D022142:Secretory Vesicles / Q000378:metabolism*; D013696:Temperature
D005456:Fluorescent Dyes; C049925:fura-2-am; D002118:Calcium; D016257:Fura-2