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Numerous terminal branches of motor nerves each end on individual component muscle fibres making up their motor unit. Each ramification terminates in a neuromuscular junction comprising pre- and postsynaptic membranes separated by the synaptic cleft. The neurotransmitter acetylcholine is released in response to depolarisation from presynaptic vesicles as discrete quanta. Each such event elicits a postsynaptic miniature endplate potential. This probabilistic release process follows a Poisson distribution whose probability increases with presynaptic depolarisation. The released acetylcholine diffuses across the synaptic gap to access postsynaptic ACh receptors, proteins each comprising five transmembrane subunits surrounding a central pore. Two of these (α) subunits each include an ACh binding site. ACh binding to both sites causes pore opening, permitting Na+ and K+ permeation, generating endplate currents demonstrable under voltage clamp. The resulting endplate potentials trigger a propagated action potential in the innervated muscle fibre when the resulting depolarisation attains the Na+ channel threshold.
The association between membrane excitation and alterations in membrane electrical impedance, its dependence on extracellular Na+ and the accompanying transmembrane Na+ fluxes measurable by isotope tracer methods, gave rise to the Na+ hypothesis for the action potential. Here, suprathreshold depolarising stimulation increases the voltage-dependent Na+ membrane conductance. The latter in turn initiates a regenerative cycle of membrane depolarisation and further channel opening, culminating in the action potential upstroke phase. Subsequent action-potential recovery to the resting potential then follows a voltage-dependent Na+ channel inactivation and more gradual K+ channel opening. This hypothesis was tested by voltage-clamp experiments determining the ionic currents required to drive depolarising membrane-potential steps in cephalopod giant axons from the resting to varying test levels. These revealed Na+ and K+ conductances whose voltage-dependences and kinetic properties could be incorporated into a successful mathematical reconstruction of the timecourse and properties of experimentally observed propagating action potentials.