Differences in nitric oxide production: a comparison of retinal ganglion cells and retinal glial cells cultured under hypoxic conditions.
The aim of this study was to compare the effects of hypoxia on nitric oxide synthase (NOS) expression and the production of NO between isolated retinal ganglion cells (RGCs) and retinal glial cells. Reverse transcription-polymerase chain reaction (RT-PCR) was employed to examine the presence of neuronal NOS mRNA, inducible NOS mRNA, and endothelial NOS mRNAs in the two cell types. RGCs and retinal glial cells were separately cultured under hypoxic (10% O(2)) or control (20% O(2)) conditions. Changes in NOS-mRNA expression were quantified by real-time PCR, and nitrite in the medium was measured up to 96 h of culture. The effects of non-NOS- and iNOS-selective inhibitors on hypoxia-induced release of nitrite in the culture medium were evaluated. RT-PCR revealed the presence of three types of NOSs in the two types of cultured cells. Hypoxic culture conditions significantly changed the expression of all NOS mRNAs in retinal glial cells but not in RGCs. NO production showed significant changes corresponding to those of NOS mRNAs in retinal glial cells but not in RGCs, and both NOS inhibitors significantly reduced hypoxia-induced nitrite release in retinal glial cells. Retinal glial cells but not RGCs may be the major source of NO under hypoxic conditions.
Kashiwagi
Kenji
K
Department of Ophthalmology, University of Yamanashi Faculty of Medicine, 1110 Shimokato, Tamaho, Yamanashi, 409-3898, Japan. kenjik@res.yamanashi-med.ac.jp
Iizuka
Yoko
Y
Mochizuki
Seiichi
S
Tsumamoto
Yuichi
Y
Mishima
Hiromu K
HK
Araie
Makoto
M
Suzuki
Yasuyuki
Y
Tsukahara
Shigeo
S
D000818:Animals; D000831:Animals, Newborn; D048429:Cell Size / Q000502:physiology; D002478:Cells, Cultured; D004791:Enzyme Inhibitors / Q000494:pharmacology; D002534:Hypoxia, Brain / Q000201:enzymology* / Q000503:physiopathology; D009457:Neuroglia / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D009569:Nitric Oxide / Q000096:biosynthesis*; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D012164:Retinal Diseases / Q000201:enzymology* / Q000503:physiopathology; D012165:Retinal Ganglion Cells / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*
D004791:Enzyme Inhibitors; D012333:RNA, Messenger; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase