Long-term impairment of subependymal repopulation following damage by ionizing irradiation

Abstract

In the mammalian brain, the subependyma (SE) contains stem cells capable of producing neurons and glia. In normal brain these stem cells are responsible, in part, for maintaining the morphologic and functional integrity of the SE; what role the cells of the SE play in brain injury has not yet been elucidated. The present study was designed to determine the long-term regenerative potential of the rat SE after significant depletion of stem cells. Ionizing irradiation was used to deplete cells of the SE and subsequent cellular responses were quantified using immunohistochemical analyses on formalin-fixed, paraffin-embedded tissues. A histomorphometric approach was used to quantify total cell number, number of proliferating cells, number of immature neurons, astrocytes, and undifferentiated components of the SE. Because there are no markers specific for stem cells, we used a repopulation assay as an indirect measure of stem cell response after injury. Our data showed clear radiation dose-dependencies in our quantitative endpoints, implying that there was progressively more stem cell damage with increasing radiation dose. Repopulation of the SE in terms of total cell number, number of proliferating cells and numbers of immature neurons was impaired in a dose-dependent fashion up to 180 days after treatment. These data suggest that after irradiation, surviving stem cells are unable to regenerate the SE. This inability to regenerate after stem cell damage/depletion could have important implications with respect to the normal function of the SE and the function of the SE after brain injury.