In vitro injury model for oligodendrocytes: development, injury, and recovery.
In this study we investigated the effects of severe hypothermia (cryoinjury) on oligodendrocyte (OL) cell marker expression and morphological features. We used a chemically defined cell culture medium, glial development medium (GDM), which favored the optimal expression of the OL phenotype in CG4 cells. Experiments using CG4 cells cultured in 2% serum or in GDM were conducted in parallel. After severe hypothermia, cells were reanimated at 37 degrees C and 4.5% CO(2) and cultured in either GDM or in medium supplemented with 2% serum. In either medium, around 70% of the total number of cells detached within 2 to 4 hours following reanimation. Oligodendroglial markers such as A2B5, O4, Tf, ferritin, tubulin, and MBP were examined by double and triple immunofluorescence. All of these markers except MBP re-appeared at different times during the recovery period for up to 48 hours. Glial fibrillary acidic protein (GFAP) and heat shock protein 60 (HSP-60) were used as injury markers. The presence of serum induced HSP-60 expression, while GDM did not. All CG4 cells expressed HSP-60 in response to hypothermia independently of the cell culture medium used. Cryoinjury induced a spectrum of morphological changes in CG4 cells. The expression of OL specific markers was also influenced by hypothermia. Moreover both, serum and cryoinjury induced the expression of HSP-60 that colocalized with OL and myelin markers. The expression of GFAP by injured cells but not by normal cells corroborated the state of injury of CG4 cells.
De Los Monteros
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AE
Mental Retardation Research Center, Neuropsychiatric Institute, Department of Neurobiology UCLA School of Medicine, 760 Westwood Plaza, Room 78-177 NPI, Los Angeles, CA 90095-1759, USA. aespinosa@mednet.ucla.edu
Zhao
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PM
De Vellis
J
J
D000818:Animals; D015415:Biomarkers / Q000032:analysis; D002478:Cells, Cultured; D018834:Chaperonin 60 / Q000378:metabolism; D003470:Culture Media; D004195:Disease Models, Animal; D005615:Freezing; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D007035:Hypothermia / Q000503:physiopathology*; D009836:Oligodendroglia / Q000502:physiology*
D015415:Biomarkers; D018834:Chaperonin 60; D003470:Culture Media; D005904:Glial Fibrillary Acidic Protein