title: Characterization of GABA- and glycine-induced currents of solitary rodent retinal ganglion cells in culture.
abstract: Ganglion cells were fluorescently labeled, dissociated from 7- to 11-day-old rodent retinas, and placed in tissue culture. Whole-cell recordings with patch electrodes were obtained from solitary cells lacking processes, which permitted a high-quality space clamp. Both GABA (1-200 microM) and glycine (10-300 microM) produced large increases in membrane conductance in virtually every ganglion cell tested, including ganglion cells from different size classes in both rats and mice. Taurine evoked responses similar to those of glycine, but considerably greater concentrations of taurine (150-300 microM) were necessary to observe any effect. Since 20 microM GABA produced approximately the same response as 100 microM glycine, the effects of these two concentrations were compared under various conditions. When recording with chloride distributed equally across the membrane, the reversal potential of the agonist-induced currents was approximately 0 mV. When the internal chloride was reduced by substitution with aspartate, the reversal potential shifted in a negative direction by about 42 mV, indicating that the current was carried mainly by chloride ions. Strychnine (1-5 microM) completely and reversibly blocked the actions of glycine (100 microM) but not those of GABA (20 microM); however, higher concentrations of strychnine (20 microM) nearly totally inhibited the current elicited by GABA (20 microM). The responses to glycine (100 microM) were not affected by bicuculline methiodide (20 microM) or picrotoxinin (20 microM). In contrast, bicuculline methiodide (10 microM) and picrotoxinin (10 microM) reversibly blocked the current evoked by GABA (20 microM); d-tubocurarine (100 microM) only slightly decreased the response to GABA (20 microM). The antagonists were effective over a wide range of holding potentials (-90 mV to +30 mV). The responses to a steady application of both GABA and glycine decayed in a few seconds when recorded under conditions of both symmetric and asymmetric chloride across the membrane. During this decay the current and conductance decreased simultaneously, reflecting receptor desensitization rather than a change in the driving force for chloride caused by agonist-induced ionic fluxes. The time-course of desensitization was usually described by a single exponential with time constants for GABA (20 microM) and glycine (100 microM) of 4.0 +/- 1.6 s and 4.4 +/- 1.9 s (mean +/- S.D.), respectively.(ABSTRACT TRUNCATED AT 400 WORDS)
authors.affiliation: Department of Neurology, Children's Hospital, Boston, MA 02115.
authors.forename: D L
authors.initials: DL
authors.lastname: Tauck
authors.forename: M P
authors.initials: MP
authors.lastname: Frosch
authors.forename: S A
authors.initials: SA
authors.lastname: Lipton
mesh_terms: D000818:Animals; D001640:Bicuculline / Q000494:pharmacology; D002478:Cells, Cultured; D005998:Glycine / Q000378:metabolism / Q000494:pharmacology*; D008564:Membrane Potentials / Q000187:drug effects; D051379:Mice; D008810:Mice, Inbred C57BL; D010852:Picrotoxin / Q000494:pharmacology; D051381:Rats; D012160:Retina / Q000502:physiology*; D012165:Retinal Ganglion Cells / Q000187:drug effects / Q000502:physiology*; D013331:Strychnine / Q000494:pharmacology; D013654:Taurine / Q000494:pharmacology; D005680:gamma-Aminobutyric Acid / Q000378:metabolism / Q000494:pharmacology*
chemical_list: D001640:Bicuculline; D005998:Glycine; D010852:Picrotoxin; D013331:Strychnine; D013654:Taurine; D005680:gamma-Aminobutyric Acid