NG2-expressing glia (NG2 cells, polydendrocytes) appear in the embryonic brain, expand perinatally, and persist widely throughout the gray and white matter of the mature central nervous system. We have previously reported that NG2 cells generate oligodendrocytes in both gray and white matter and a subset of protoplasmic astrocytes in the gray matter of the ventral forebrain and spinal cord. To investigate the temporal changes in NG2 cell fate, we generated NG2creER™BAC transgenic mice, in which tamoxifen-inducible Cre is expressed in NG2 cells. Cre induction at embryonic day 16.5, postnatal day (P) 2, P30 and P60 in mice that were double transgenic for NG2creER™BAC and the Cre reporter revealed that NG2 cells in the postnatal brain generate only NG2 cells or oligodendrocytes, whereas NG2 cells in the embryonic brain generate protoplasmic astrocytes in the gray matter of the ventral forebrain in addition to oligodendrocytes and NG2 cells. Analysis of cell clusters from single NG2 cells revealed that more than 80% of the NG2 cells in the P2 brain give rise to clusters consisting exclusively of oligodendrocytes, whereas the majority of the NG2 cells in the P60 brain generate clusters that contain only NG2 cells or a mixture of oligodendrocytes and NG2 cells. Furthermore, live cell imaging of single NG2 cells from early postnatal brain slices revealed that NG2 cells initially divide symmetrically to produce two daughter NG2 cells and that differentiation into oligodendrocytes occurred after 2-3 days.

Cells that express the NG2 proteoglycan (NG2+ cells) constitute a large cell population in the adult mammalian central nervous system (CNS). They give rise to mature oligodendrocytes in culture and are thus considered to be oligodendrocyte progenitor cells (OPCs). They proliferate in response to a variety of insults to the CNS, but their ability to differentiate into oligodendrocytes in vivo has not been established. We used bromodeoxyuridine (BrdU) to trace the fate of NG2+ cells that proliferated in response to a chemically induced demyelinating lesion in the adult rat spinal cord. Cells that were proliferating 24 hr after lesioning were labeled by a single injection of BrdU, and their antigenic phenotype was examined at various times up to 28 days post‐lesioning (28 dpl). Initially, at 2 dpl, NG2+/BrdU+ cells were found almost exclusively at the periphery of the lesion. At 7 dpl, the number of NG2+/BrdU+ cells increased in the lesion center and decreased from the surrounding areas. The number of NG2+/BrdU+ cells inside the lesion further decreased with time, concomitant with progression of remyelination and appearance of BrdU+ mature oligodendrocytes. Double labeling with ³H‐thymidine and BrdU combined with NG2 immunohistochemistry showed that some NG2+ cells in the lesion had undergone at least two rounds of cell division. These observations strongly suggest that NG2+/BrdU+ cells that appeared in response to the demyelinating insult gave rise to mature remyelinating oligodendrocytes, providing an in vivo evidence for the differentiation of NG2+ cells into oligodendrocytes.

Oligodendrocyte precursor cells (OPCs) retain their proliferative/differentiation capacity throughout life. This study uses in vivo and ex vivo imaging to show a specific temporal window between OPC division and oligodendrocyte differentiation in the postnatal mouse brain that is modulated by the microenvironment. The latency between OPC division and differentiation is shortened by myelin damage, while sensory deprivation reduces the survival of divided OPCs undergoing differentiation. Oligodendrocytes in the mammalian brain are continuously generated from NG2 cells throughout postnatal life. However, it is unclear when the decision is made for NG2 cells to self-renew or differentiate into oligodendrocytes after cell division. Using a combination of in vivo and ex vivo imaging and fate analysis of proliferated NG2 cells in fixed tissue, we demonstrate that in the postnatal developing mouse brain, the majority of divided NG2 cells differentiate into oligodendrocytes during a critical age-specific temporal window of 3–8 d. Notably, within this time period, damage to myelin and oligodendrocytes accelerated oligodendrocyte differentiation from divided cells, and whisker removal decreased the survival of divided cells in the deprived somatosensory cortex. These findings indicate that during the critical temporal window of plasticity, the fate of divided NG2 cells is sensitive to modulation by external signals.