Local cell proliferation Upon enucleation in Direct Retinal Brain Targets in the Visual system of the Adult Mouse

In this study we used incorporation of the DNA synthesis marker 5-bromo-2’-deoxyuridine or BrdU to visualize cell proliferation in the visual system of the adult mouse as a response to monocular enucleation. We detected new BrdU-labeled cells in different subcortical retinal target regions and we established a specific time frame in which this cell proliferation occurred. By performing immunofluorescent double stainings for BrdU and different vascular (glucose transporter type 1, collagen type IV), glial (thymosin β4, glial fibrillary acidic protein) and neuronal (Neuronal Nuclei, doublecortin) markers, we identified these proliferating cells as activated microglia. Additional immunohistochemical stainings for thymosin β4 and glial fibrillary acidic protein also revealed reactive astrocytes in the different retinorecipient nuclei and allowed us to delineate a time frame for microglial and astroglial activation. A PCR array experiment further showed increased levels of cytokines, chemokines, growth factors and enzymes that play an important role in microglial-astroglial communication during the glial activation process in response to the deafferentation.