Murine encephalitogenic lymphoid cells induce nitric oxide synthase in primary astrocytes.
A cytokine-inducible form of nitric oxide synthase (iNOS), capable of producing large quantities of nitric oxide (NO), can be induced in many cell types. We demonstrate that conditioned medium from encephalitogenic myelin basic protein-sensitized lymphoid cells (MBP-CM) induces the expression of iNOS in primary cultures of murine astrocytes in a time- and concentration-dependent manner. iNOS mRNA was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) as early as 3 h post-exposure. Accumulation of nitrite into the astrocyte culture medium, an indirect measure of NO, was measurable 3 h post-exposure, plateaued at 24 h, and was prevented by the simultaneous administration of the NOS inhibitors, L-N(G)-nitroarginine methyl ester, N(G)-nitro-L-arginine or aminoguanidine. Astrocyte expression of iNOS protein, detected by immunohistochemistry and immunoprecipitation/Western blot, was prevented by inhibitors of RNA or protein metabolism, consistent with its dependence on de novo protein synthesis.
Hewett
S J
SJ
Department of Neurology, Center for the Study of Nervous System Injury, Washington University School of Medicine, St. Louis, MO 63110, USA.
Misko
T P
TP
Keeling
R M
RM
Behrens
M M
MM
Choi
D W
DW
Cross
A H
AH
D000818:Animals; D001253:Astrocytes / Q000201:enzymology*; D002478:Cells, Cultured; D004681:Encephalomyelitis, Autoimmune, Experimental / Q000276:immunology*; D004790:Enzyme Induction; D005260:Female; D007158:Immunologic Techniques; D008198:Lymph Nodes / Q000166:cytology / Q000502:physiology*; D051379:Mice; D008815:Mice, Inbred Strains; D004676:Myelin Basic Protein / Q000276:immunology; D019001:Nitric Oxide Synthase / Q000096:biosynthesis* / Q000235:genetics; D016133:Polymerase Chain Reaction; D014158:Transcription, Genetic
D004676:Myelin Basic Protein; D019001:Nitric Oxide Synthase