Expression of a dominant negative IFN-gammareceptor on mouse oligodendrocytes.
The interferon-gamma (IFN-gamma) receptor is expressed by all nucleated cells, and binding of its cognate ligand, IFN-gamma, induces a wide variety of biological functions. Transgenic mice expressing a dominant negative IFN-gamma receptor 1 (IFN-gammaR1DeltaIC) on oligodendrocytes under control of the myelin proteolipid protein promoter are described. The mRNA encoding the transgene was only detected in the nervous system and protein expression was confirmed by immunohistochemistry. Transgenic receptor expression does not alter myelination and the mice exhibited no clinically apparent phenotype. Consistent with the restricted nervous system expression of the transgene, no alterations in peripheral immune responses were detected. Flow cytometric analysis demonstrated constitutive expression of both the IFN-gammaR1DeltaIC transgene and the endogenous IFN-gamma receptor 2 at high levels on oligodendrocytes derived from the transgenic mice. These oligodendrocytes also exhibited decreased STAT1 phosphorylation in response to IFN-gamma, confirming dominant negative transgene function. Transgenic mice in which oligodendrocytes have a diminished ability to respond to IFN-gamma showed delayed virus clearance from oligodendroglia compared with wild-type mice. This model will allow evaluation of oligodendrocyte responses to this critical cytokine during CNS inflammation.
González
John M
JM
Department of Neurology, University of Southern California, Keck School of Medicine, Los Angeles, California 90033, USA.
Bergmann
Cornelia C
CC
Fuss
Babette
B
Hinton
David R
DR
Kangas
Cindy
C
Macklin
Wendy B
WB
Stohlman
Stephen A
SA
D000818:Animals; D015153:Blotting, Western; D002490:Central Nervous System / Q000166:cytology; D004268:DNA-Binding Proteins / Q000378:metabolism; D005434:Flow Cytometry; D007109:Immunity / Q000502:physiology; D007150:Immunohistochemistry; D007371:Interferon-gamma / Q000378:metabolism*; D051379:Mice; D018991:Myelin Proteolipid Protein / Q000378:metabolism; D009836:Oligodendroglia / Q000378:metabolism*; D010766:Phosphorylation; D010957:Plasmids / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D017471:Receptors, Interferon / Q000096:biosynthesis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D050794:STAT1 Transcription Factor; D015398:Signal Transduction / Q000187:drug effects; D013116:Spinal Cord / Q000187:drug effects / Q000378:metabolism; D015534:Trans-Activators / Q000378:metabolism; D019076:Transgenes / Q000235:genetics
D004268:DNA-Binding Proteins; D018991:Myelin Proteolipid Protein; D012333:RNA, Messenger; D017471:Receptors, Interferon; D050794:STAT1 Transcription Factor; C494080:Stat1 protein, mouse; D015534:Trans-Activators; D007371:Interferon-gamma