title: Inhibition of reactive gliosis attenuates excitotoxicity-mediated death of retinal ganglion cells.
abstract: Reactive gliosis is a hallmark of many retinal neurodegenerative conditions, including glaucoma. Although a majority of studies to date have concentrated on reactive gliosis in the optic nerve head, very few studies have been initiated to investigate the role of reactive gliosis in the retina. We have previously shown that reactive glial cells synthesize elevated levels of proteases, and these proteases, in turn, promote the death of retinal ganglion cells (RGCs). In this investigation, we have used two glial toxins to inhibit reactive gliosis and have evaluated their effect on protease-mediated death of RGCs. Kainic acid was injected into the vitreous humor of C57BL/6 mice to induce reactive gliosis and death of RGCs. C57BL/6 mice were also treated with glial toxins, alpha-aminoadipic acid (AAA) or Neurostatin, along with KA. Reactive gliosis was assessed by immunostaining of retinal cross sections and retinal flat-mounts with glial fibrillary acidic protein (GFAP) and vimentin antibodies. Apoptotic cell death was assessed by TUNEL assays. Loss of RGCs was determined by immunostaining of flat-mounted retinas with Brn3a antibodies. Proteolytic activities of matrix metalloproteinase-9 (MMP-9), tissue plasminogen activator (tPA), and urokinase plasminogen activator (uPA) were assessed by zymography assays. GFAP-immunoreactivity indicated that KA induced reactive gliosis in both retinal astrocytes and in Muller cells. AAA alone or in combination with KA decreased GFAP and vimentin-immunoreactivity in Mϋller cells, but not in astrocytes. In addition AAA failed to decrease KA-mediated protease levels and apoptotic death of RGCs. In contrast, Neurostatin either alone or in combination with KA, decreased reactive gliosis in both astrocytes and Mϋller cells. Furthermore, Neurostatin decreased protease levels and prevented apoptotic death of RGCs. Our findings, for the first time, indicate that inhibition of reactive gliosis decreases protease levels in the retina, prevents apoptotic death of retinal neurons, and provides substantial neuroprotection.
authors.affiliation: Eye Research Institute of Oakland University, Rochester, Minnesota, United States of America.
authors.forename: Bhagyalaxmi S
authors.initials: BS
authors.lastname: Ganesh
authors.forename: Shravan K
authors.initials: SK
authors.lastname: Chintala
mesh_terms: D015074:2-Aminoadipic Acid / Q000627:therapeutic use; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D005911:Gliosis / Q000139:chemically induced / Q000188:drug therapy* / Q000378:metabolism; D006028:Glycosphingolipids / Q000627:therapeutic use*; D007150:Immunohistochemistry; D020287:In Situ Nick-End Labeling; D066298:In Vitro Techniques; D007608:Kainic Acid / Q000627:therapeutic use / Q000633:toxicity; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D012160:Retina / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D012165:Retinal Ganglion Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D010959:Tissue Plasminogen Activator / Q000378:metabolism; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism
chemical_list: D006028:Glycosphingolipids; C116059:neurostatin; D015074:2-Aminoadipic Acid; D010959:Tissue Plasminogen Activator; D014568:Urokinase-Type Plasminogen Activator; D020780:Matrix Metalloproteinase 9; D007608:Kainic Acid