Effects of calcium on the dynamic process of transmitter release which generates either skew- or bell-MEPPs

Miniature endplate potential (MEPP) amplitudes, MEPP frequencies and ratios of skew:bell-MEPPs were determined as well as synaptic vesicle diameters and densities at the mouse diaphragm neuromuscular endplate during exposure to elevated calcium concentrations. Additions of external Ca2+ had variable effects on MEPP frequencies and percentages of skew-MEPPs, regardless of concentrations used (1-25 mM). Nevertheless, changes in MEPP amplitudes were most sensitive (4-fold decrease) to low value increases of Ca2+. Changes in MEPP frequencies produced by an increase in Ca2+ were very sensitive to initial frequencies as well as the initial calcium concentration. An increase in Ca2+ usually increased MEPP frequency (providing skew-MEPPs were measured). Changes in the percentage of skew-MEPPs were extremely variable (4-90%) and these changes depended on initial frequencies, initial skew- to bell-MEPP ratios and age of the mouse. With a change in Ca2+ concentration, synaptic vesicle diameters and densities remained constant during changes in MEPP frequencies and large changes in the skew:bell-MEPP ratios; and, vesicle numbers were sometimes slightly increased. Because of the wide range in MEPP frequencies and amplitudes, this study demonstrates that the effect of various treatments should be evaluated on identified endplates and that analyses of randomly selected endplates must consider the large variability between endplates. These results show that the skew-MEPP class must not be ignored in studies of spontaneous MEPP release, and that initial frequencies and age of the mouse are also important in evaluating changes in skew-MEPP to bell-MEPP ratios. The rapid changes in skew- to bell-MEPP classes indicate that MEPP class and size are determined at the moment of release by the state of the release process as proposed by Kriebel et al. (1990). Because changes in calcium concentration can immediately alter the ratio of skew- to bell-MEPPs we conclude that the release process has two states to generate the two classes of MEPPs, and that the release process is very sensitive to conditions so that states are easily changed. We propose that the release process meters transmitter in subunit amounts to form both classes of MEPPS and that the calcium ions modulate the process.