Comparison of vesicular volume and quantal size in bovine chromaffin cells

Electrochemical measurements of vesicular content released were compared with the morphometric measurements of vesicular size in bovine chromaffin cells. Cross-sectional vesicular diameters were determined from electron micrographs. Two methods were used to determine the frequency histograms of “true” vesicular diameters (i.e. diameters of the vesicles in the equatorial plane): (i) “peeling off” method [Coupland R. E. (1968) Nature 217, 384–388], and (ii) summation of individual probabilities of “true” vesicular diameters. Quantal size was estimated from the area under the spontaneous current spike detected electrochemically. The frequency histograms of “true” vesicular diameters are found to be skewed (thus not well described by a Gaussian function) irrespective of the method used to calculate them, as are the frequency histograms of the cube roots of the quantal sizes. Furthermore, we also find that the frequency histograms of electrochemical measurements (the cube roots of quantal sizes) have lower skews and coefficients of variation than those of morphometric measurements (“true” vesicular diameters), with discrepancy being especially pronounced for noradrenaline-secreting cells. Such a difference in both coefficients of variation and skews suggests that the intravesicular catecholamine concentration is not uniform, but that it is lower for vesicles of larger size. In conclusion a variety of factors—vesicular volume, vesicular surface area to volume ratio, binding capacity of chromogranin and/or ATP, likely determines the amount of catecholamine stored in the vesicle.