Yamashita_1994_Brain.Res.Dev.Brain.Res_82_95

It has been shown in previous studies that a subpopulation of neurons in monolayer cultures prepared from immature embryonic chicken retina acquired a series of functional properties which characterized them as GABAergic amacrine cells after 1 week in vitro. In the present study, we demonstrate that immature precursors of these cells were already identifiable by morphological criteria after 2 days in vitro (DIV). Using the whole cell patch-clamp technique we have studied the time-course of the expression of voltage-dependent and of glutamate and GABA receptor-associated conductances in these identified retinal interneurons developing in vitro. Recordings after 2 DIV revealed a very homogeneous pattern of membrane conductances. In all cells tested, whole cell responses to depolarizing voltage steps consisted solely of a sustained outward potassium current and 100% of the cells responded to the glutamate receptor agonist kainic acid (KA) and to GABA. Fast activating inward sodium currents first appeared after 3 DIV, whereas a transient component of outward potassium currents was not detectable before day 4 in vitro. N-Methyl-D-aspartate (NMDA)-evoked currents were first observed at 3 DIV in the GABAergic neurons. Only 1 day later they were found in all of the GABAergic neurons. Expression of responses to quisqualic acid (QU) started at 3 DIV, but remained restricted to a subpopulation of the GABAergic cells even at later stages (59% at 4 DIV, 63% at 6-9 DIV). Antagonistic effects of QU on KA responses, however, were detectable in all cells tested, independent of the developmental stage and the presence of QU-evoked currents.