Decrease of quantal size and quantal content during tetanic stimulation detected by focal recording.

End-plate potentials and miniature end-plate potentials were recorded focally (i.e., over a limited area of the end-plate with several or possibly only one active zone) in a cutaneous pectoris from neuromuscular junction during a prolonged (1-6 min) tetanic (20-100 Hz) nerve stimulation. End-plate potential amplitudes decreased and became more variable with prolonged stimulation. Synaptic depression thus occurs even when synaptic output is low, if release is evoked from only a few active zones, suggesting that there is little if any vesicular replenishment between the active zones. The probability density function of the end-plate potential amplitudes has been obtained using the Parzen estimate with a Gaussian weighting function, to reduce the number of end-plate potentials needed for the same accuracy. Quantal size of the end-plate potentials was estimated from the slope of the best fitted line to the prominent and apparently equidistant peaks of probability density functions or from the spectrogram of the probability density function of end-plate potentials. Quantal contents were initially (+/- S.D.):5.7 +/- 2.9, ranged from 2 to 12, and in all cases examined (n = 11) decreased with prolonged tetanic stimulation. The rates of the decrease of end-plate potentials amplitudes (and quantal contents) from different segments of the same nerve terminal were often different, even when they were initially comparable. This suggests that some active zones or some areas of the end-plates become depleted much faster than others. Quantal sizes of the nerve evoked and the spontaneously released quanta were generally similar at low frequencies of stimulation (0.5-2 Hz). Both decreased with high frequency stimulation, but the decrease of the quantal sizes of nerve evoked quanta was usually more pronounced. At different loci of the same end-plate the contribution of lower quantal size to the synaptic depression varied widely (from < 5% to > 80%). In conclusion lower quantal size can contribute significantly to synaptic depression. At uneven decrease of quantal sizes over the whole nerve terminal helps to explain both aspects of synaptic depression (lower synaptic efficacy and greater variability of quantal responses.