Possible involvement of Janus kinase Jak2 in interferon-gamma induction of nitric oxide synthase in rat glial cells.
To clarify the induction pathway of inducible nitric oxide (NO) synthase in the brain, we examined the effects of interferon-gamma and lipopolysaccharide on the induction of inducible NO synthase in glial cells cultured from neonatal rats, compared to those in the macrophage cell line RAW264.7 which was derived from Abelson leukemia virus-induced BALB/c lymphocytic lymphoma. NO synthase activity (NO2- accumulation) and 130 kDa protein of inducible NO synthase were induced 24 h after treatment with interferon-gamma or lipopolysaccharide in both glial cells and RAW264.7 macrophages. These induction activities were inhibited by a tyrosine kinase inhibitor, herbimycin A. Immunoprecipitation assay using antibodies against Janus kinases, and the signal transducer and activator of transcription-1 (STAT1), revealed that interferon-gamma induced tyrosine phosphorylation of the just another kinase-2 (Jak2) and STAT1 alpha but did not induced the phosphorylation of Jak1, the non-receptor tyrosine kinase-2 (Tyk2) and STAT1 beta. Tyrosine phosphorylation of Jak2 and STAT1 alpha induced by interferon-gamma was also inhibited by herbimycin A, while lipopolysaccharide did not induce any tyrosine phosphorylation of Janus kinases and STAT1 at all. These results suggest that the interferon-gamma-induced inducible NO synthase induction involves activation of Jak2-STAT1 alpha pathway in both glial cells and macrophages.
Kitamura
Y
Y
Department of Neurobiology, Kyoto Pharmaceutical University, Japan.
Takahashi
H
H
Nomura
Y
Y
Taniguchi
T
T
D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D004790:Enzyme Induction / Q000187:drug effects; D007371:Interferon-gamma / Q000494:pharmacology; D053614:Janus Kinase 2; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000187:drug effects / Q000201:enzymology; D051379:Mice; D008807:Mice, Inbred BALB C; D009457:Neuroglia / Q000187:drug effects / Q000201:enzymology*; D019001:Nitric Oxide Synthase / Q000378:metabolism*; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D011518:Proto-Oncogene Proteins; D011817:Rabbits; D051381:Rats; D017471:Receptors, Interferon / Q000302:isolation & purification / Q000378:metabolism*; D011994:Recombinant Proteins; D015398:Signal Transduction / Q000502:physiology*
D008070:Lipopolysaccharides; D011518:Proto-Oncogene Proteins; D017471:Receptors, Interferon; D011994:Recombinant Proteins; D007371:Interferon-gamma; D019001:Nitric Oxide Synthase; D011505:Protein-Tyrosine Kinases; C507926:Jak2 protein, mouse; C507927:Jak2 protein, rat; D053614:Janus Kinase 2