In situ localization of nitric oxide synthase and direct evidence of NO production in rat retinal ganglion cells

Brain Research
Yuichi TsumamotoHiromu K Mishima

Abstract

The expression of isoforms of nitric oxide synthase (NOS), enzymes responsible for NO production, and the synthesis of nitric oxide (NO) in rat retinal ganglion cells (RGCs) during synaptogenesis for various phases of the pre- and postnatal developmental periods were investigated. The retinas from prenatal, lactating, young, and adult rats were fixed in paraformaldehyde. The cryosections or paraformaldehyde-fixed ganglion cells purified from rat pups were immunostained for constitutive isoforms of NOS (n and eNOS) and observed with a confocal laser scanning microscope. Synthesis of NO in the RGCs was achieved by in vitro stimulation with glutamate. The intracellular NO levels were measured in real time using diaminofluorescein-2 diacetate, a fluorescence indicator of NO. Immunohistochemical analysis revealed nNOS and eNOS expressed in retinal ganglion cells during the first 2 postnatal weeks. Cultured RGCs also expressed nNOS and eNOS in vitro. Intracellular NO levels in cultured RGCs showed spontaneous fluctuation during a 20-min observation. The presence of both a non-specific NOS inhibitor, L-NAME, and a specific nNOS inhibitor, 7-NI, significantly inhibited (P<0.001) the increase of intracellular NO 6 and 8 min after the in...Continue Reading

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Citations

Sep 28, 2005·Japanese Journal of Ophthalmology·Ryotaro GotoYukitaka Uji
Sep 2, 2010·Purinergic Signalling·Narinobu Harada
Jul 24, 2003·Neuroscience Letters·Hideo HoshiTeruya Ohtsuka
Dec 7, 2013·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Ryan E TookerJozsef Vigh
May 30, 2009·Journal of Neuroscience Research·Thomas J GioveWilliam D Eldred
Dec 3, 2011·Brain Research·Alex H VielmaOliver Schmachtenberg
Jan 28, 2004·Journal of Pharmacological Sciences·Hiroshi KatsukiAkinori Akaike
Jan 21, 2006·World Journal of Gastroenterology : WJG·Hong-Mei WangYi-Ling Wang
Feb 22, 2005·Experimental Eye Research·Enri BordaLeonor Sterin-Borda

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