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Vesicular release of neurotransmitters: converting amperometric measurements into size, dynamics and energetics of initial fusion pores.

Amperometric currents displaying a pre-spike feature (PSF) may be treated so as to lead to precise information about initial fusion pores, viz., about the crucial event initiating neurotransmitter vesicular release in neurons and medullary glands. However, amperometric data alone are not self-sufficient, so their full exploitation requires external calibration to solve the inverse problem. For this purpose we resorted to patch-clamp measurements published in the literature on chromaffin cells. Reported pore radii were thus used to evaluate the diffusion rate of neurotransmitter cations in the partially altered matrix located near the fusion pore entrance. This allowed an independent determination of each initial fusion pore radius giving rise to a single PSF event. The statistical distribution of the radii thus obtained provided for the first time an experimental access to the potential energy well governing the thermodynamics of such systems. The shape of the corresponding potential energy well strongly suggested that, after their creation, initial fusion pores are essentially controlled by the usual physicochemical laws describing pores formed in bilayer lipidic biological membranes, i.e., they have an essentially lipidic nature.