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The Journal of Neuroscience, October 1, 2000, 20(19):7290-7296
Stimulation-Evoked Increases in Cytosolic [Ca2+] in
Mouse Motor Nerve Terminals Are Limited by Mitochondrial Uptake and Are
Temperature-Dependent
Gavriel
David and
Ellen F.
Barrett
Department of Physiology and Biophysics, University of Miami School
of Medicine, Miami, Florida 33101
Increases in cytosolic [Ca2+] evoked by trains
of action potentials (20-100 Hz) were recorded from mouse and lizard
motor nerve terminals filled with a low-affinity fluorescent indicator,
Oregon Green BAPTA 5N. In mouse terminals at near-physiological
temperatures (30-38°C), trains of action potentials at 25-100 Hz
elicited increases in cytosolic [Ca2+] that
stabilized at plateau levels that increased with stimulation frequency.
Depolarization of mitochondria with carbonylcyanide m-chlorophenylhydrazone (CCCP) or antimycin A1
caused cytosolic [Ca2+] to rise to much higher
levels during stimulation. Thus, mitochondrial Ca2+
uptake contributes importantly to limiting the rise of cytosolic [Ca2+] during repetitive stimulation.
In mouse terminals, the stimulation-induced increase in cytosolic
[Ca2+] was highly temperature-dependent over the
range 18-38°C, with greater increases at lower temperatures. At the
lower temperatures, application of CCCP continued to depolarize
mitochondria but produced a much smaller increase in the cytosolic
[Ca2+] transient evoked by repetitive stimulation.
This result suggests that the larger amplitude of the
stimulation-induced cytosolic [Ca2+] transient at
lower temperatures was attributable in part to reduced
mitochondrial Ca2+ uptake.
In contrast, the stimulation-induced increases in cytosolic
[Ca2+] measured in lizard motor terminals showed
little or no temperature-dependence over the range 18-33°C.
Key words:
mitochondria; presynaptic terminal; motor nerve terminal; calcium indicator dyes; calcium sequestration; neuromuscular junction; lizard; temperature
Copyright © 2000 Society for Neuroscience 0270-6474/00/20197290-07$05.00/0
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