Direct evidence for electrical coupling among rat supraoptic nucleus neurons

Abstract

Transfer of the fluorescent dye, Lucifer yellow (LY), from an intracellularly injected neuron to one or more other neurons is accepted as indirect evidence of electrotonic interactions among such dye coupled cells. Direct evidence requires that at least two coupled cells be recorded from simultaneously and such evidence in the CNS has been gained only for hippocampal pyramidal neurons. Since interpretations of the functional significance of dye coupling among magnocellular neuroendocrine cells depend upon its relation to electrical coupling, we sought to obtain direct evidence for electrotonic interactions in such neurons. Over 150 pairs of supraoptic nucleus (SON) neurons in hypothalamic slices were recorded from intracellularly using one LY and one potassium acetate electrode in each instance. Of these, 9 pairs were studied in sufficient detail to determine that they were electrically coupled. Most of the remaining pairs were determined not to be coupled. In each coupled pair of cells, membrane voltage changes due to spontaneously occurring or current evoked action potentials, as well as current evoked hyperpolarizations, in one cell were reflected in similar, though attenuated changes in the other cell. All of these changes occurred simultaneously in the two neurons. Spontaneously arising postsynaptic potentials in the two cells were temporally uncorrelated. In each case that electrical coupling was observed, dye coupling resulted from LY injection. Coupling ratios ranged from 0.05 to 0.2. Capacitative coupling between the recording electrodes as an artifact was ruled out since cells in the same tissue penetration as the coupled cell showed no responses to membrane voltage changes in the primary cell; no responses were seen with the second electrode placed extracellularly or in the medium; and similar coupling potentials were also seen when one cell was recorded without a second electrode present. We conclude that electrical coupling exists among magnocellular neurons of the SON and that the incidence of dye coupling is a reasonable estimate of the incidence of electrical coupling. These electrotonic interactions probably play important roles in the coordination of firing among magnocellular neurosecretory neurons.