Calcium channels and GABA receptors in the early embryonic chick retina

The properties of calcium channels were studied at the period of neurogenesis in the early embryonic chick retina. The whole neural retina was isolated from embryonic day 3 (E3) chick and loaded with a Ca(2+)-sensitive fluorescent dye (Fura-2). The retinal cells were depolarized by puff application of high-K+ solutions. Increases in intracellular Ca2+ concentrations were evoked by the depolarization through calcium channels. The type of calcium channel was identified as L-type by the sensitivity to dihydropyridines. The Ca2+ response was completely blocked by 10 microM nifedipine, whereas it was remarkably enhanced by 5 microM Bay K 8644. Then we sought a factor to activate the calcium channel and found that GABA could activate it by membrane depolarization at the E3 chick retina. Puff application of 100 microM GABA raised intracellular Ca2+ concentrations, and this Ca2+ response to GABA was also sensitive to the two dihydropyridines. Intracellular potential recordings verified clear depolarization by bath-applied 100 microM GABA. The Ca2+ response to GABA was mediated by GABAA receptors, since the GABA response was blocked by 10 microM bicuculline or 50 microM picrotoxin, and mimicked by muscimol but not by baclofen. Neither glutamate, kainate, nor glycine evoked any Ca2+ response. We conclude that L-type calcium channels and GABAA receptors are already expressed before differentiation of retinal cells and synapse formation in the chick retina. A possibility is proposed that GABA might act as a trophic factor by activating L-type calcium channels via GABAA receptors during the early period of retinal neurogenesis.