title: Estimation of presynaptic calcium currents and endogenous calcium buffers at the frog neuromuscular junction with two different calcium fluorescent dyes.
abstract: At the frog neuromuscular junction, under physiological conditions, the direct measurement of calcium currents and of the concentration of intracellular calcium buffers-which determine the kinetics of calcium concentration and neurotransmitter release from the nerve terminal-has hitherto been technically impossible. With the aim of quantifying both Ca(2+) currents and the intracellular calcium buffers, we measured fluorescence signals from nerve terminals loaded with the low-affinity calcium dye Magnesium Green or the high-affinity dye Oregon Green BAPTA-1, simultaneously with microelectrode recordings of nerve-action potentials and end-plate currents. The action-potential-induced fluorescence signals in the nerve terminals developed much more slowly than the postsynaptic response. To clarify the reasons for this observation and to define a spatiotemporal profile of intracellular calcium and of the concentration of mobile and fixed calcium buffers, mathematical modeling was employed. The best approximations of the experimental calcium transients for both calcium dyes were obtained when the calcium current had an amplitude of 1.6 ± 0.08 pA and a half-decay time of 1.2 ± 0.06 ms, and when the concentrations of mobile and fixed calcium buffers were 250 ± 13 μM and 8 ± 0.4 mM, respectively. High concentrations of endogenous buffers define the time course of calcium transients after an action potential in the axoplasm, and may modify synaptic plasticity.
authors.affiliation: Laboratory of the Biophysics of Synaptic Processes, Kazan Scientific Centre, Kazan Institute of Biochemistry and Biophysics, Russian Academy of Sciences Kazan, Russia ; Open Laboratory of Neuropharmacology, Kazan Federal University Kazan, Russia ; Department of Radiophotonics and Microwave Technologies, Kazan National Research Technical University named after A. N. Tupolev Kazan, Russia.
authors.forename: Dmitry
authors.initials: D
authors.lastname: Samigullin
authors.affiliation: Laboratory of the Biophysics of Synaptic Processes, Kazan Scientific Centre, Kazan Institute of Biochemistry and Biophysics, Russian Academy of Sciences Kazan, Russia.
authors.forename: Nijaz
authors.initials: N
authors.lastname: Fatikhov
authors.affiliation: Laboratory of the Biophysics of Synaptic Processes, Kazan Scientific Centre, Kazan Institute of Biochemistry and Biophysics, Russian Academy of Sciences Kazan, Russia ; Open Laboratory of Neuropharmacology, Kazan Federal University Kazan, Russia.
authors.forename: Eduard
authors.initials: E
authors.lastname: Khaziev
authors.affiliation: Laboratory of the Biophysics of Synaptic Processes, Kazan Scientific Centre, Kazan Institute of Biochemistry and Biophysics, Russian Academy of Sciences Kazan, Russia ; Department of Neurobiology and Radioelectronics, Kazan Federal University Kazan, Russia.
authors.forename: Andrey
authors.initials: A
authors.lastname: Skorinkin
authors.affiliation: Laboratory of the Biophysics of Synaptic Processes, Kazan Scientific Centre, Kazan Institute of Biochemistry and Biophysics, Russian Academy of Sciences Kazan, Russia ; Open Laboratory of Neuropharmacology, Kazan Federal University Kazan, Russia ; Department of Medical and Biological Physics, Kazan State Medical University Kazan, Russia.
authors.forename: Eugeny
authors.initials: E
authors.lastname: Nikolsky
authors.affiliation: Laboratory of the Biophysics of Synaptic Processes, Kazan Scientific Centre, Kazan Institute of Biochemistry and Biophysics, Russian Academy of Sciences Kazan, Russia ; Open Laboratory of Neuropharmacology, Kazan Federal University Kazan, Russia.
authors.forename: Ellya
authors.initials: E
authors.lastname: Bukharaeva
keywords: endogenous calcium buffers; inward calcium current; mathematical modeling; neuromuscular junction; presynaptic calcium transient