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J Neurophysiol 85: 287-294, 2001;
0022-3077/01 $5.00
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The Journal of Neurophysiology Vol. 85 No. 1 January 2001, pp. 287-294
Copyright ©2001 by the American Physiological Society

Intraterminal Ca2+ and Spontaneous Transmitter Release at the Frog Neuromuscular Junction

J. K. Angleson1 and W. J. Betz2

 1Department of Biological Sciences, University of Denver, Denver 80208; and  2Department of Physiology and Biophysics, University of Colorado Medical School, Denver, Colorado 80262

Angleson, J. K. and W. J. Betz. Intraterminal Ca2+ and Spontaneous Transmitter Release at the Frog Neuromuscular Junction. J. Neurophysiol. 85: 287-294, 2001. We investigated the relationship between intraterminal Ca2+ concentration ([Ca2+]i) and the frequency of miniature end plate potentials (MEPPs) at the frog neuromuscular junction by use of ratiometric imaging of fura-2-loaded nerve terminals and intracellular recording of MEPPs. Elevation of extracellular [KCl] over the range of 2-20 mM resulted in increases in [Ca2+]i and MEPP frequency. Loading terminals with the fast and slow Ca2+-buffers bis-(o-aminophenoxy)-N,N,N',N'-tetraacetic acid-acetoxymethyl (BAPTA-AM) and EGTA-AM resulted in equivalent reductions in the KCl-dependent increases in MEPP frequency. The [Ca2+]i dependence of MEPP frequency determined by elevation of [Ca2+]i due to application of 0.1-10 µM ionomycin was similar to that determined when [Ca2+]i was raised by increasing extracellular KCl. Measurements in 10 mM extracellular KCl revealed that application of the phorbol ester phorbol 12 myristate 13-acetetate (PMA) caused an increase in MEPP frequency while the inactive analogue, 4alpha -PMA, did not. PMA application also caused an increase in [Ca2+]i. The relationship between [Ca2+]i and MEPP frequency in PMA was the same as was determined by the other methods of raising [Ca2+]i. Under all conditions tested, our data revealed a low [Ca2+]i threshold for activation of transmitter release and are consistent with a Kd for [Ca2+]i on the order of 1 µM.




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