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The Journal of Neuroscience, November 15, 2000, 20(22):8377-8383
The Readily Releasable Pool of Vesicles in Chromaffin Cells Is
Replenished in a Temperature-Dependent Manner and Transiently Overfills
at 37°C
Vera
Dinkelacker,
Thomas
Voets,
Erwin
Neher, and
Tobias
Moser
Department of Membrane Biophysics, Max-Planck-Institute for
Biophysical Chemistry, D-37077 Göttingen, Germany
Maturation of exocytic vesicles to the release-ready state is
regulated by several factors, including intracellular calcium concentration ([Ca2+]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
[Ca2+]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°C, respectively) yielding a Q10 of
2.3. The changes of the Ca2+ signal between the
different temperatures could not account for the differences in
recovery kinetics. At 32 and 37°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 [Ca2+]int,
hence with a large difference between prestimulus and poststimulus [Ca2+]int. In cells with higher basal
[Ca2+]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 Ca2+ transient.
Key words:
exocytosis; chromaffin cells; capacitance; vesicle pools; refilling; temperature; calcium
Copyright © 2000 Society for Neuroscience 0270-6474/00/20228377-07$05.00/0
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