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(Investigative Ophthalmology and Visual Science. 2000;41:4169-4174.)
© 2000 by The Association for Research in Vision and Ophthalmology, Inc.

RGC Death in Mice after Optic Nerve Crush Injury: Oxidative Stress and Neuroprotection

Hana Levkovitch–Verbin1, Cathy Harris–Cerruti2, Yoram Groner2, Larry A. Wheeler3, Michal Schwartz4,5 and Eti Yoles4,5

1 From the Goldschleger Eye Institute, Sheba Medical Center, Sackler School of Medicine, Tel-Aviv University, Israel; 2 Departments of Molecular Genetics and 3 Neurobiology, The Weizmann Institute of Science, Rehovot, Israel; and the 4 Department of Biological Sciences, Allergen, Irvine, California.

PURPOSE. To establish a method for morphometric analysis of retrogradely labeled retinal ganglion cells (RGCs) of the mouse retina, to be used for the study of molecular aspects of RGC survival and neuroprotection in this model; to evaluate the effect of overexpression of Cu-Zn-superoxide dismutase (CuZnSOD) on RGC survival after severe crush injury to the optic nerve, and to assess the effect of the {alpha}2-adrenoreceptor agonist brimonidine, recently shown to be neuroprotective, on RGC survival.

METHODS. A severe crush injury was inflicted unilaterally in the orbital portion of the optic nerves of wild-type and transgenic (Tg–SOD) mice expressing three to four times more human CuZnSOD than the wild type. In each mouse all RGCs were labeled 72 hours before crush injury by stereotactic injection of the neurotracer dye FluoroGold (Fluorochrome, Denver, CO) into the superior colliculus. Survival of RGCs was then assessed morphometrically, with and without systemic injection of brimonidine.

RESULTS. Two weeks after crush injury, the number of surviving RGCs was significantly lower in the Tg-SOD mice (596.6 ± 71.9 cells/mm2) than in the wild-type control mice (863.5 ± 68 cells/mm2). There was no difference between the numbers of surviving RGCs in the uninjured retinas of the two strains (3708 ± 231.3 cells/mm2 and 3904 ± 120 cells/mm2, respectively). Systemic injections of brimonidine significantly reduced cell death in the Tg-SOD mice, but not in the wild type.

CONCLUSIONS. Overexpression of CuZnSOD accelerates RGC death after optic nerve injury in mice. Activation of the {alpha}2-adrenoreceptor pathway by brimonidine enhances survival of RGCs in an in vivo transgenic model of excessive oxidative stress.




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