Tsugihisa Sasaki1, Akimichi Kaneko. 1. Department of Ophthalmology, Kanazawa University School of Medicine, Kanazawa, Ishikawa, Japan. sasatsug@w5.dion.ne.jp
Abstract
PURPOSE: To analyze the mechanism of hypoxia-induced changes of the intracellular Ca(2+) concentration ([Ca(2+)](i)) in retinal ganglion cells (RGCs). METHODS: Fluo-3 was applied to the cut edge of the optic nerve of 6-week-old rats. The retina was sliced, and the Ca images were captured. A hypoxic condition was created by superfusing the retinal slice with an oxygen/glucose-deprived solution. RESULTS: The retrograde staining method filled the RGCs selectively. Fifteen minutes of hypoxic conditions induced an increase in [Ca(2+)](i) in the RGCs (Delta0.13 +/- 0.03, n = 23). Application of 60 microM DL-2-amino-5-phosphonovaleric acid partially blocked the hypoxia-induced [Ca(2+)](i) increase in dendrites (Delta0.03 +/- 0.02, n = 4, P < 0.05) but not in the somata (Delta0.12 +/- 0.02, n = 9). The RGC dendrites showed a further increase in [Ca(2+)](i) after being switched back to an oxygenated solution (Delta0.14 +/- 0.04, n = 4). Neither 6-cyano-7-nitroquinoxaline-2,3-dione disodium, DL: -threo-beta-benzyloxyaspartate, nifedipine, nor bepridil inhibited the hypoxia-induced [Ca(2+)](i) increase. A Ca(2+)-free superfusion prevented the hypoxia-induced [Ca(2+)](i) increase in the somata (Delta0.07 +/- 0.02, n = 5, P < 0.05) but not in the dendrites (Delta0.16 +/- 0.005, n = 4). CONCLUSIONS: The mechanism of the hypoxia-induced increase in [Ca(2+)](i) differs between somata and dendrites. The N-methyl-D-aspartate channel of dendrites seems to be the main route of Ca(2+) influx. Copyright (c) Japanese Ophthalmological Society 2007.
PURPOSE: To analyze the mechanism of hypoxia-induced changes of the intracellular Ca(2+) concentration ([Ca(2+)](i)) in retinal ganglion cells (RGCs). METHODS:Fluo-3 was applied to the cut edge of the optic nerve of 6-week-old rats. The retina was sliced, and the Ca images were captured. A hypoxic condition was created by superfusing the retinal slice with an oxygen/glucose-deprived solution. RESULTS: The retrograde staining method filled the RGCs selectively. Fifteen minutes of hypoxic conditions induced an increase in [Ca(2+)](i) in the RGCs (Delta0.13 +/- 0.03, n = 23). Application of 60 microM DL-2-amino-5-phosphonovaleric acid partially blocked the hypoxia-induced [Ca(2+)](i) increase in dendrites (Delta0.03 +/- 0.02, n = 4, P < 0.05) but not in the somata (Delta0.12 +/- 0.02, n = 9). The RGC dendrites showed a further increase in [Ca(2+)](i) after being switched back to an oxygenated solution (Delta0.14 +/- 0.04, n = 4). Neither 6-cyano-7-nitroquinoxaline-2,3-dione disodium, DL: -threo-beta-benzyloxyaspartate, nifedipine, nor bepridil inhibited the hypoxia-induced [Ca(2+)](i) increase. A Ca(2+)-free superfusion prevented the hypoxia-induced [Ca(2+)](i) increase in the somata (Delta0.07 +/- 0.02, n = 5, P < 0.05) but not in the dendrites (Delta0.16 +/- 0.005, n = 4). CONCLUSIONS: The mechanism of the hypoxia-induced increase in [Ca(2+)](i) differs between somata and dendrites. The N-methyl-D-aspartate channel of dendrites seems to be the main route of Ca(2+) influx. Copyright (c) Japanese Ophthalmological Society 2007.