Metallothioneins are upregulated in symptomatic mice with astrocyte-targeted expression of tumor necrosis factor-alpha.
Transgenic mice expressing TNF-alpha under the regulatory control of the GFAP gene promoter (GFAP-TNFalpha mice) exhibit a unique, late-onset chronic-progressive neurological disorder with meningoencephalomyelitis, neurodegeneration, and demyelination with paralysis. Here we show that the metallothionein-I + II (MT-I + II) isoforms were dramatically upregulated in the brain of symptomatic but not presymptomatic GFAP-TNFalpha mice despite TNF-alpha expression being present in both cases. In situ hybridization analysis for MT-I RNA and radioimmunoassay results for MT-I + II protein revealed that the induction was observed in the cerebellum but not in other brain areas. Increased MT-I RNA levels occurred in the Purkinje and granular neuronal layers of the cerebellum but also in the molecular layer. Reactive astrocytes, activated rod-like microglia, and macrophages, but not the infiltrating lymphocytes, were identified as the cellular sources of the MT-I + II proteins. In situ hybridization for MT-III RNA revealed a modest increase in the white matter of the cerebellum, which was confirmed by immunocytochemistry. MT-III immunoreactivity was present in cells which were mainly round or amoeboid monocytes/macrophages. The pattern of expression of the different MT isoforms in the GFAP-TNFalpha mice differed substantially from that described previously in GFAP-IL6 mice, demonstrating unique effects associated with the expression of each cytokine. The results suggest that the MT expression in the CNS reflects the inflammatory response and associated damage rather than a direct role of the TNF-alpha in their regulation and support a major role of these proteins during CNS injury.
Carrasco
J
J
Departamento de Biología Celular, de Fisiología, y de Inmunología, Unidad de Fisiología Animal, Bellaterra, Barcelona, 08193, Spain.
Giralt
M
M
Penkowa
M
M
Stalder
A K
AK
Campbell
I L
IL
Hidalgo
J
J
D000818:Animals; D001253:Astrocytes / Q000166:cytology / Q000378:metabolism*; D001933:Brain Stem / Q000378:metabolism / Q000473:pathology; D002531:Cerebellum / Q000378:metabolism / Q000473:pathology; D003711:Demyelinating Diseases / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D004027:Diencephalon / Q000378:metabolism / Q000473:pathology; D018450:Disease Progression; D018390:Gene Targeting; D005904:Glial Fibrillary Acidic Protein / Q000235:genetics; D008668:Metallothionein / Q000378:metabolism*; D000090023:Metallothionein 3; D051379:Mice; D008822:Mice, Transgenic; D009419:Nerve Tissue Proteins / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics; D016548:Prosencephalon / Q000378:metabolism / Q000473:pathology; D020033:Protein Isoforms / Q000096:biosynthesis / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis* / Q000235:genetics; D015854:Up-Regulation
D005904:Glial Fibrillary Acidic Protein; D000090023:Metallothionein 3; D009419:Nerve Tissue Proteins; D020033:Protein Isoforms; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D008668:Metallothionein