Illumination partly reverses the postsynaptic blockade of the frog neuromuscular junction by the styryl pyridinium dye RH414

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Abstract
The styryl pyridinium dye RH414, which we have used recently for optical monitoring of synaptic vesicle membrane trafficking in frog motor nerve terminals, reversibly inhibited contractions in the frog nerve-muscle preparations used for those studies. We report here the results of experiments into the basis of this inhibition and how the blocking effects can be modulated by incident light, by using conventional intracellular recording and iontophoretic techniques. Bath application of 5-42.5 microM RH414 blocked nerve-evoked twitches in frog cutaneus pectoris muscles, although subthreshold endplate potentials and miniature endplate potentials could still be recorded. In the presence of the dye, illumination of the recording area with light from a mercury arc lamp over a wide range of wavelengths (340-560 nm) potentiated the amplitude of endplate potentials, miniature endplate potentials, and depolarizations resulting from iontophoretic application of acetylcholine. The magnitude of both the light-induced disinhibition of endplate and iontophoretic potentials increased with the intensity of the illumination and developed exponentially with a time constant of several hundred milliseconds. Both the dye-induced inhibition and the light-induced disinhibition disappeared after washing in dye-free physiological saline. Muscle fibre resting membrane potential, input conductance and miniature endplate potential frequency, however, were not affected by these manipulations. These data are consistent with a specific, curare-like interaction of the dye with acetylcholine receptors which can be modulated by light.