NG2 cells are not a major source of reactive astrocytes after neocortical stab wound injury

Abstract

NG2 cells are an abundant glial cell type in the adult brain. They are distinct from astrocytes, mature oligodendrocytes, and microglia. NG2 cells generate oligodendrocytes and a subpopulation of protoplasmic astrocytes in the ventral forebrain during development. To determine whether NG2 cells generate reactive astrocytes in the lesioned brain, stab wound injury was created in adult NG2creBAC:ZEG double transgenic mice, in which enhanced green fluorescent protein (EGFP) is expressed in NG2 cells and their progeny, and the phenotype of the EGFP(+) cells was analyzed at 10 and 30 days post lesion (dpl). The majority (>90%) of the reactive astrocytes surrounding the lesion that expressed glial fibrillary acidic protein (GFAP) lacked EGFP expression, and conversely the majority (>90%) of EGFP(+) cells were GFAP-negative. However, 8% of EGFP(+) cells co-expressed GFAP at 10 dpl. Most of these EGFP(+) GFAP(+) cells were morphologically distinct from hypertrophic reactive astrocytes and exhibited weak GFAP expression. NG2 was detected in a fraction of the EGFP(+) GFAP(+) cells found at 10 dpl. By 30 dpl the number of EGFP(+) GFAP(+) cells had decreased more than four-fold from 10 dpl. A similar transient appearance of EGFP(+) GFAP(+) cells with simple morphology was observed in NG2creER™:ZEG double transgenic mice in which EGFP expression had been induced in NG2 cells prior to injury. NG2 cell-specific deletion of the oligodendrocyte lineage transcription factor Olig2 using NG2creER™:Olig2(fl/fl) :ZEG triple transgenic mice did not increase the number of EGFP(+) reactive astrocytes. These findings suggest that NG2 cells are not a major source of reactive astrocytes in the neocortex.