The probability of quantal secretion along visualized terminal branches at amphibian (Bufo marinus) neuromuscular synapses

Abstract

The number of quanta secreted from selected sites along terminal branches at toad (Bufo marinus) neuromuscular junctions was determined. Terminal branches were visualized by prior staining with the fluorescent dye, 3-3 Diethyloxadicarbocyanine iodide (DiOC2(5)); neither impulse conduction nor quantal release were affected by DiOC2(5) at concentrations less than 10 microM. The evoked quantal release recorded with an extracellular micro-electrode (me) at different sites along the length of terminal branches was determined in an external calcium concentration, [Ca]o, of 0.35-0.45 mM. For short branches (40-80 microns), me remained approximately constant for over 60% of the branches; for the rest, me declined approximately exponentially with an average length constant of 17 +/- 2 microns (mean +/- S.E. of mean). For both medium (81-120 microns) and long branches (121-160 microns), me declined in nearly all cases approximately exponentially with length constants of 39 +/- 5 and 54 +/- 8 microns respectively. These changes in me were observed at synapses having a wide range of terminal branching patterns. Some DiOC2(5)-stained branches possessed discontinuous cholinesterase staining. In general, me declined along these branches in the same way as along DiOC2(5)-stained branches with continuous cholinesterase staining. It is suggested that because of the decline in me along most medium and long terminal branches, many release sites have a very low probability for secretion in low [Ca]o. Release sites near the point of nerve entry, which have a relatively high probability, therefore make the main contribution to secretion recorded with an intracellular micro-electrode. As a consequence, transmitter secretion from the whole terminal does not fluctuate from impulse to impulse as much as expected if there were a large number of release sites, each with a low probability of secretion. Transmitter secretion then follows binomial rather than Poisson statistics.