Regulation of Nerve Growth Factor Release by Nitric Oxide through Cyclic GMP Pathway in Cortical Glial Cells
- Huabao Xiong1,
- Kiyofumi Yamada2,
- Hussam Jourdi1,
- Meiko Kawamura1,
- Nobuyuki Takei1,
- Daiken Han2,
- Toshitaka Nabeshima2 and
- Hiroyuki Nawa1
- 1Department of Molecular Neurobiology, Brain Research Institute, Niigata University, Niigata, Japan (H.X., H.J., M.K., N.T., H.N.); and2Department of Neuropsychopharmacology and Hospital Pharmacy, Nagoya University School of Medicine, Nagoya, Japan (K.Y., D.H., T.N.)
Abstract
In the present study, we found thatS-nitroso-N-acetyl-dl-penicillamine, a spontaneous nitric oxide (NO) generator, dose-dependently inhibited basal nerve growth factor (NGF) release from mixed glial cells. To elucidate the function of endogenous NO in the regulation of NGF release, the mixed glial cells were stimulated with lipopolysaccharide (LPS) or LPS plus interferon-γ (IFNγ). The results showed that LPS alone induced NGF release and moderate NO production. However, costimulation with LPS plus IFNγ greatly enhanced NO production but significantly suppressed LPS-induced NGF release. WhenNG-monomethyl-l-arginine, an NOS inhibitor, was added to the culture, the suppression of NGF release by IFNγ was significantly reduced. Quantitative reverse transcription-polymerase chain reaction demonstratedS-nitroso-N-acetyl-dl-penicillamine was also able to inhibit the LPS-induced NGF mRNA expression. To understand the different contributions of astroglia and microglia to this phenomenon, both cell types were purified. We found purified astroglia produced high amounts of NGF but low amounts of NO. However, purified microglia produced a large amount of NO but very low amounts of NGF after stimulation with LPS or LPS plus IFNγ. Our data also indicated the second messenger cyclic GMP, but not cyclic AMP, was able to inhibit basal NGF release. In vivo experiments confirmed that NGF protein level was significantly enhanced in rats treated withl-Nω-nitro-arginine methyl ester and in endothelial NO synthase mutant mice. Taken together, we conclude NO derived mainly from microglia down-regulates NGF release from astroglia at the transcriptional level by stimulating cyclic GMP pathway.
Footnotes
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Send reprint requests to: Huabao Xiong, M.D., Ph.D., Department of Molecular Neurobiology, Brain Research Institute, Niigata University, 1-757, Asahimachi-dori, Niigata 951-8585, Japan. E-mail:xionghbl{at}bri.niigata-u.ac.jp
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This study was supported by the Japan Society for the Promotion of Science (RFTF-96L00203).
- Abbreviations:
- NGF
- nerve growth factor
- EIA
- enzyme immunoassay
- CNS
- central nervous system
- cGMP
- cyclic GMP
- cAMP
- cyclic AMP
- IFN
- interferon
- GFAP
- antiglial fibrillary acidic protein
- LPS
- lipopolysaccharide
- iNOS
- inducible NOS
- nNOS
- neuronal NOS
- eNOS
- endothelial NOS
- NAME
- Nω-nitro-arginine methyl ester
- TBST
- Tween 20-containing Tris-buffered saline
- NMMA
- NG-monomethyl-l-arginine
- NO
- nitric oxide
- NOS
- nitric oxide synthase
- PCR
- polymerase chain reaction
- SNAP
- S-nitroso-N-acetyl-dl-penicillamine
- TNF
- tumor necrosis factor
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- Received January 26, 1999.
- Accepted May 18, 1999.
- The American Society for Pharmacology and Experimental Therapeutics
