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J Neurophysiol 87: 1816-1823, 2002;
0022-3077/02 $5.00
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The Journal of Neurophysiology Vol. 87 No. 4 April 2002, pp. 1816-1823
Copyright ©2002 by the American Physiological Society

Ca2+-Dependent Ca2+ Clearance Via Mitochondrial Uptake and Plasmalemmal Extrusion in Frog Motor Nerve Terminals

S. Suzuki,1 M. Osanai,1 N. Mitsumoto,2 T. Akita,1 K. Narita,3 H. Kijima,4 and K. Kuba1

 1Department of Physiology, School of Medicine, Nagoya University, Nagoya 466-8550;  2Department of Physiology, Saga Medical School, Saga 849-8501;  3Department of Physiology, Kawasaki Medical School, Kurashiki 701-0192; and  4Department of Physics, School of Science, Nagoya University, Nagoya 464-8602, Japan

Suzuki, S., M. Osanai, N. Mitsumoto, T. Akita, K. Narita, H. Kijima, and K. Kuba. Ca2+-Dependent Ca2+ Clearance Via Mitochondrial Uptake and Plasmalemmal Extrusion in Frog Motor Nerve Terminals. J. Neurophysiol. 87: 1816-1823, 2002. Ca2+ clearance in frog motor nerve terminals was studied by fluorometry of Ca2+ indicators. Rises in intracellular Ca2+ ([Ca2+]i) in nerve terminals induced by tetanic nerve stimulation (100 Hz, 100 or 200 stimuli: Ca2+ transient) reached a peak or plateau within 6-20 stimuli and decayed at least in three phases with the time constants of 82-87 ms (81-85%), a few seconds (11-12%), and several tens of seconds (less than a few percentage). Blocking both Na/Ca exchangers and Ca2+ pumps at the cell membrane by external Li+ and high external pH (9.0), respectively, increased the time constants of the initial and second decay components with no change in their magnitudes. By contrast, similar effects by Li+ alone, but not by high alkaline alone, were seen only on 200 stimuli-induced Ca2+ transients. Blocking Ca2+ pumps at Ca2+ stores by thapsigargin did not affect 100 stimuli-induced Ca2+ transients but increased the initial decay time constant of 200 stimuli-induced Ca2+ transients with no change in other parameters. Inhibiting mitochondrial Ca2+ uptake by carbonyl cyanide m-chlorophenylhydrazone markedly increased the initial and second decay time constants of 100 stimuli-induced Ca2+ transients and the amplitudes of the second and the slowest components. Plotting the slopes of the decay of 100 stimuli-induced Ca2+ transients against [Ca2+]i yielded the supralinear [Ca2+]i dependence of Ca2+ efflux out of the cytosol. Blocking Ca2+ extrusion or mitochondrial Ca2+ uptake significantly reduced this [Ca2+]i-dependent Ca2+ efflux. Thus Ca2+-dependent mitochondrial Ca2+ uptake and plasmalemmal Ca2+ extrusion clear out a small Ca2+ load in frog motor nerve terminals, while thapsigargin-sensitive Ca2+ pump boosts the clearance of a heavy Ca2+ load. Furthermore, the activity of plasmalemmal Ca2+ pump and Na/Ca exchanger is complementary to each other with the slight predominance of the latter.




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