Inmaculada Rite

Reactive gliosis is the universal reaction to brain injury, but the precise origin and subsequent fate of the glial cells reacting to injury are unknown. Astrocytes react to injury by hypertrophy and up-regulation of the glial-fibrillary acidic protein (GFAP). Whereas mature astrocytes do not normally divide, a subpopulation of the reactive GFAP(+) cells does so, prompting the question of whether the proliferating GFAP(+) cells arise from endogenous glial progenitors or from mature astrocytes that start to proliferate in response to brain injury. Here we show by genetic fate mapping and cell type-specific viral targeting that quiescent astrocytes start to proliferate after stab wound injury and contribute to the reactive gliosis and proliferating GFAP(+) cells. These proliferating astrocytes remain within their lineage in vivo, while a more favorable environment in vitro revealed their multipotency and capacity for self-renewal. Conversely, progenitors present in the adult mouse cerebral cortex labeled by NG2 or the receptor for the platelet-derived growth factor (PDGFRalpha) did not form neurospheres after (or before) brain injury. Taken together, the first fate-mapping analysis of astrocytes in the adult mouse cerebral cortex shows that some astrocytes acquire stem cell properties after injury and hence may provide a promising cell type to initiate repair after brain injury.

We have evaluated the possibility that changes in the vascular system may constitute a contributing factor for the death of nigral dopaminergic neurons in Parkinson's disease. Thus, we have employed intranigral injections of vascular endothelial growth factor (VEGF), the most potent inducer of blood-brain barrier (BBB) permeability. A single dose of 1 mug of VEGF, chosen from a dose-response study, highly disrupted the BBB in the ventral mesencephalon in a time-dependent manner. A strong regional correlation between BBB disruption and loss of tyrosine hydroxylase-positive neurons was evident. Moreover, Fluoro-Jade B labelling showed the presence of dying neurons in the substantia nigra in response to VEGF injection. High number of TUNEL-positive nuclei was observed in this area along with activation of caspase 3 within nigral dopaminergic neurons. Analysis of the glial population demonstrated a strong inflammatory response and activation of astroglia in response to BBB disruption. We conclude that disruption of the BBB may be a causative factor for degeneration of nigral dopaminergic neurons.

Aquaporin-4 (AQP4) is the most abundant aquaporin in the brain and it is widely accepted that this AQP is solely expressed by astrocytes and ependymal cells. AQP4 is particularly enriched in plasma membranes of ependymal cells and astrocyte membrane domains facing blood vessels and pia. AQP4 has gained much attraction due to its involvement in the physiopathology of brain edema, a major cause of death in humans. Consequently, it is of paramount importance to ascertain the phenotypic nature of AQP4 mRNA-expressing cells in the CNS before attempting future clinical studies aimed at minimizing the development of brain edema. We have used intranigral injections of lipopolysaccharide (LPS), a potent immunostimulant that causes disruption of the blood brain barrier, vasogenic edema, loss of reactive astrocytes and activation of microglial cells. These LPS-induced features are ideal for testing the possibility that reactive microglial cells express AQP4 in the adult brain. We have studied AQP4 at the mRNA and protein level. We provide strong evidence that reactive microglial cells highly express AQP4 mRNA and protein in response to LPS injections.