Interaction of calcium-permeable non-N-methyl-D-aspartate receptor channels with voltage-activated potassium and calcium currents in rat retinal ganglion cells in vitro.
Calcium-permeable non-N-methyl-D-aspartate receptor channels are now characterized in much detail, but still little is known about the consequences of Ca2+ influx through these channels in specific neuron types. We are interested in the role of Ca2+-permeable non-N-methyl-D-aspartate receptor channels during differentiation of retinal ganglion cells. However, in view of the conflicting data on the relative Ca2+ permeability of non-N-methyl-D-aspartate receptor channels in these neurons, a more systematic evaluation of permeation properties of different Na+ substitutes was necessary before proceeding with the main goal of the present study evaluating the effects of non-N-methyl-D-aspartate receptor activation on repetitive firing and voltage-activated K+ and Ca2+ conductances. Retinal ganglion cells were dissociated from the rat retina on postnatal day 5. They were selected by vital anti-Thy-1 immunostaining and repetitive firing behaviour and submitted to patch-clamp recording in the whole-cell configuration. Non-N-methyl-D-aspartate receptor channels were activated by application of amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid or kainate. It was found that they were essentially impermeable to N-methyl-D-glucamine (P(NMDG)/P(Cs)<0.02), but not to choline (P(choline)/P(Cs)=0.24) and tetramethylammonium (P(TMA)/P(Cs)=0.23). When using N-methyl-D-glucamine as a substitute for Na+ to obtain bi-ionic conditions P(Ca)/P(Cs) varied between 0.08 to 1.40. Linear current voltage relation or little outward rectification corresponded to a low Ca2+ permeability (P(Ca)/P(Cs)=0.14). In about one third of the cells kainate-induced currents showed inward rectification and non-N-methyl-D-aspartate receptor agonists induced a substantially higher Ca2+ influx (P(Ca)/P(Cs)=0.64). Activation of non-N-methyl-D-aspartate receptors by kainate profoundly altered the repetitive discharge of retinal ganglion cells. In contrast to the continuously firing controls, cells generated only a few spikes at the beginning of a steady depolarization after kainate exposure. Among the candidates regulating the firing behaviour of retinal ganglion cells voltage-activated Ca2+ and K+ conductances were tested for their sensitivity to kainate application. It was found that even short conditioning pulses of kainate decreased the peak amplitudes of both voltage-activated K+ and voltage-activated Ca2+ currents. Only the latter effect required extracellular Ca2+ and was antagonized by increasing the intracellular Ca2+ buffering strength. Thus, suppression of calcium currents was induced by a non-N-methyl-D-aspartate receptor-mediated rise of the intracellular calcium concentration. The reduction of K+ currents did not depend on extracellular calcium and was insensitive to experimental manipulation of intracellular Ca2+ buffer strength. The interaction between Ca2+-permeable non-N-methyl-D-aspartate receptor channels and voltage-activated Ca2+ and K+ currents may represent an important regulatory mechanism to control the repetitive firing of developing retinal ganglion cells.
Taschenberger
H
H
Arbeitsgruppe Entwicklungsneurobiologie, Institut für Physiologie der Charité, Humboldt-Universität Zu Berlin, Germany.
Grantyn
R
R
D000818:Animals; D000831:Animals, Newborn; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000378:metabolism*; D002478:Cells, Cultured; D004594:Electrophysiology; D015640:Ion Channel Gating / Q000502:physiology; D008564:Membrane Potentials / Q000502:physiology; D018408:Patch-Clamp Techniques; D015221:Potassium Channels / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D016194:Receptors, N-Methyl-D-Aspartate / Q000378:metabolism*; D012165:Retinal Ganglion Cells / Q000378:metabolism*
D015220:Calcium Channels; D015221:Potassium Channels; D016194:Receptors, N-Methyl-D-Aspartate; D002118:Calcium