Neuronal Isoform of Nitric Oxide Synthase is Expressed at Low Levels in Human Retina

Overview

Journal Cell Mol Neurobiol

Publisher Springer

Specialties Cell Biology
Molecular Biology
Neurology

Date 1996 Aug 1

PMID 8879752

Citations 3

Authors

C S Park

C Gianotti

R Park

G Krishna

Affiliations

Soon will be listed here.

Abstract

1. The expression of neuronal isoform of nitric oxide synthase (nNOS) was studied in human retinal tissues. The cDNA sequence was cloned in human retinal poly (A)+ RNA by the RT-PCR method and encompassed an open-reading frame of 4,302 bp encoding 1,434 amino acids. This sequence showed a possibility of genetic polymorphism in comparison to human brain form. 2. Restriction fragment length polymorphism (RFLP) patterns of a partial cDNA fragment suggest that there is genetic polymorphism in the neuronal form of NOS. Important differences were observed in a certain region between human retinal and brain froms. This region is a result of frame shift by the addition of three cytidines. In this study, regions from human brain (cerebellum) and skeletal muscle as well as retina were sequenced to confirm the difference in this region. The sequences from these tissues were completely identical. This indicated that genetic polymorphism of nNOS gene was due to single base substitution and not frame shift phenomenon by addition or deletion of bases. 3. The nNOS mRNA of approximately 12 kb was detected by northern blot analysis. The lower level of the expression was distinguished in comparison to those of human brain and skeletal muscle. The cDNA transiently transfected into CHO-K1 cells expressed a protein which contained a significant level of NOS activity. The size of the nNOS was found to be approximately 160 kDa by both in vitro and in vivo translation systems. This NOS was calcium dependent and the K(m) for arginine was 4.4 microM. 4. The Ca+2, L-arginine and NADPH dependency along with the inhibitory effect of N-nitro-L-arginine on NOS activity were evaluated. The finding of a constitutive from of NOS in human retina, which is calcium-NADPH dependent, gives further credence to the possible role of nitric oxide in retinal function and neuronal diseases.

Citing Articles

Stem cell activation in skeletal muscle regeneration.

Fu X, Wang H, Hu P Cell Mol Life Sci. 2015; 72(9):1663-77.

PMID: 25572293 PMC: 4412728. DOI: 10.1007/s00018-014-1819-5.

Disease gene candidates revealed by expression profiling of retinal ganglion cell development.

Wang J, Kunzevitzky N, Dugas J, Cameron M, Barres B, Goldberg J J Neurosci. 2007; 27(32):8593-603.

PMID: 17687037 PMC: 2885852. DOI: 10.1523/JNEUROSCI.4488-06.2007.

The effect of divalent cations on bovine retinal NOS activity.

Geyer O, Podos S, Oron Y, Mittag T Mol Cell Biochem. 2000; 204(1-2):11-6.

PMID: 10718619 DOI: 10.1023/a:1007037307370.

References

Ferrendelli J . Distribution and regulation of cyclic GMP in the central nervous system. Adv Cyclic Nucleotide Res. 1978; 9:453-64. View

Mollace V, Salvemini D, ANGGARD E, Vane J . Cultured astrocytoma cells inhibit platelet aggregation by releasing a nitric oxide-like factor. Biochem Biophys Res Commun. 1990; 172(2):564-9. DOI: 10.1016/0006-291x(90)90710-5. View

Lambert L, Whitten J, Baron B, Cheng H, Doherty N, McDONALD I . Nitric oxide synthesis in the CNS endothelium and macrophages differs in its sensitivity to inhibition by arginine analogues. Life Sci. 1991; 48(1):69-75. DOI: 10.1016/0024-3205(91)90426-c. View

Gross S, Stuehr D, Aisaka K, Jaffe E, Levi R, Griffith O . Macrophage and endothelial cell nitric oxide synthesis: cell-type selective inhibition by NG-aminoarginine, NG-nitroarginine and NG-methylarginine. Biochem Biophys Res Commun. 1990; 170(1):96-103. DOI: 10.1016/0006-291x(90)91245-n. View

Krishna G, Krishnan N, FLETCHER R, Chader G . Effects of light on cyclic GMP metabolism in retinal photoreceptors. J Neurochem. 1976; 27(3):717-22. DOI: 10.1111/j.1471-4159.1976.tb10399.x. View

Venturini C, Knowles R, Palmer R, Moncada S . Synthesis of nitric oxide in the bovine retina. Biochem Biophys Res Commun. 1991; 180(2):920-5. DOI: 10.1016/s0006-291x(05)81153-2. View

Vincent S, Kimura H . Histochemical mapping of nitric oxide synthase in the rat brain. Neuroscience. 1992; 46(4):755-84. DOI: 10.1016/0306-4522(92)90184-4. View

Bredt D, Hwang P, Glatt C, Lowenstein C, Reed R, Snyder S . Cloned and expressed nitric oxide synthase structurally resembles cytochrome P-450 reductase. Nature. 1991; 351(6329):714-8. DOI: 10.1038/351714a0. View

Fujisawa H, Ogura T, Kurashima Y, Yokoyama T, Yamashita J, Esumi H . Expression of two types of nitric oxide synthase mRNA in human neuroblastoma cell lines. J Neurochem. 1994; 63(1):140-5. DOI: 10.1046/j.1471-4159.1994.63010140.x. View

10.

Baylor D . Photoreceptor signals and vision. Proctor lecture. Invest Ophthalmol Vis Sci. 1987; 28(1):34-49. View

11.

Ahmad I, Leinders-Zufall T, Kocsis J, Shepherd G, Zufall F, Barnstable C . Retinal ganglion cells express a cGMP-gated cation conductance activatable by nitric oxide donors. Neuron. 1994; 12(1):155-65. DOI: 10.1016/0896-6273(94)90160-0. View

12.

Goureau O, LEPOIVRE M, Becquet F, Courtois Y . Differential regulation of inducible nitric oxide synthase by fibroblast growth factors and transforming growth factor beta in bovine retinal pigmented epithelial cells: inverse correlation with cellular proliferation. Proc Natl Acad Sci U S A. 1993; 90(9):4276-80. PMC: 46489. DOI: 10.1073/pnas.90.9.4276. View

13.

Snyder S, Bredt D . Nitric oxide as a neuronal messenger. Trends Pharmacol Sci. 1991; 12(4):125-8. DOI: 10.1016/0165-6147(91)90526-x. View

14.

Provis J, Mitrofanis J . NADPH-diaphorase neurones of human retinae have a uniform topographical distribution. Vis Neurosci. 1990; 4(6):619-23. DOI: 10.1017/s0952523800005812. View

15.

Kristensson K, Aldskogius M, Peng Z, Olsson T, Aldskogius H, Bentivoglio M . Co-induction of neuronal interferon-gamma and nitric oxide synthase in rat motor neurons after axotomy: a role in nerve repair or death?. J Neurocytol. 1994; 23(8):453-9. DOI: 10.1007/BF01184069. View

16.

Hope B, Michael G, Knigge K, Vincent S . Neuronal NADPH diaphorase is a nitric oxide synthase. Proc Natl Acad Sci U S A. 1991; 88(7):2811-4. PMC: 51329. DOI: 10.1073/pnas.88.7.2811. View

17.

Janssens S, Shimouchi A, Quertermous T, Bloch D, Bloch K . Cloning and expression of a cDNA encoding human endothelium-derived relaxing factor/nitric oxide synthase. J Biol Chem. 1992; 267(21):14519-22. View

18.

Park C, Pardhasaradhi K, Gianotti C, Villegas E, Krishna G . Human retina expresses both constitutive and inducible isoforms of nitric oxide synthase mRNA. Biochem Biophys Res Commun. 1994; 205(1):85-91. DOI: 10.1006/bbrc.1994.2633. View

19.

Mollace V, Colasanti M, Rodino P, Massoud R, Lauro G, Nistico G . Cytokine-induced nitric oxide generation by cultured astrocytoma cells involves Ca(++)-calmodulin-independent NO-synthase. Biochem Biophys Res Commun. 1993; 191(2):327-34. DOI: 10.1006/bbrc.1993.1221. View

20.

Lyons C, Orloff G, Cunningham J . Molecular cloning and functional expression of an inducible nitric oxide synthase from a murine macrophage cell line. J Biol Chem. 1992; 267(9):6370-4. View