COMPLEMENT AND GLIAL ACTIVITY IN THE RETINOCOLLICULAR PATHWAY OF MICE USING A NOVEL MODEL OF GLAUCOMA

Purpose: Glaucoma is a leading cause of irreversible visual impairment and blindness throughout the world. C1q is responsible for axonal pruning in early ocular development and is upregulated in glaucomatous eyes of mice, non-human primates, and humans. We used an inducible mouse model of human primary open angle glaucoma with elevated intraocular pressure (IOP) to examine expression levels of C1q in the retina and superior colliculus (SC), as well as identify changes in cellular homeostasis. Methods: Anesthetized A/J mice were given a single intravitreal injection of Ad5.MYOC.Y437H (5x107 pfu), a mutant glaucoma gene, or Ad5.null control virus. Following injections, conscious IOPs were measured weekly, using a TonoLab tonometer (iCare). Mice were sacrificed at time points between 3 days and 8 weeks. Brains and retinas were harvested for immunofluorescence or immunoblotting studies. Microglia and astrocytes cells were identified using Iba1 and GFAP, respectively. All quantifications were performed using ImageJ Analysis software(NIH). Results: IOPs were significantly increased in the Ad5.MYOC.Y437H eyes (p<0.01) compared to the contralateral un-injected eye and eyes receiving Ad5.null. Clq expression was significantly upregulated in retinas receiving Ad5.MYOC.Y437H (2.69-fold±0.38, p<0.0001) compared to contralateral control retinas (0.7-fold±0.29). Clq upregulation was additionally observed in SC hemispheres receiving neural connections from injected eyes. Mice given Ad5.null vector displayed no elevation of Clq in the visual axis. Additionally, colocalization studies demonstrated significant increases of inner retinal microglia density beginning 2 weeks post injection (0.61%±0.07, p<0.001) and continuing at 4 weeks (0.87%±0.09, p<0.0001) compared to untreated retinas (0.4l%±0.03 and 0.44%±0.03, respectively). No signs of astrogliosis were detected. Conclusions: C1q is actively upregulated in the retina and SC, following mutant myocilin induced ocular hypertention, whereas adenovirus alone had no effect. An increased microglial population in the retina accompanied these changes. This suggests that microglia may sense the increased IOP and play a role in upregulating endogenous C1q. Early glaucoma pathogenesis may result from the reactivation of the ocular developmental roles of C1q and microglia, suggesting new therapeutic targets for future neuroprotective studies.