In vivo dynamics of retinal microglial activation during neurodegeneration: confocal ophthalmoscopic imaging and cell morphometry in mouse glaucoma.

摘要: Microglia, which are CNS-resident neuroimmune cells, transform their morphology and size in response to CNS damage, switching an activated state with distinct functions gene expression profiles. The roles of microglial activation health, injury disease remain incompletely understood due dynamic complex regulation changes microenvironment. Thus, it is critical non-invasively monitor analyze over time the intact organism. In vivo studies have been delayed by technical limitations tracking behavior without altering environment. This has particularly challenging during chronic neurodegeneration, where long-term must be tracked. retina, a organ amenable non-invasive live imaging, offers powerful system visualize characterize dynamics microglia disorders. This protocol outlines methods for long-term, imaging retinal microglia, using confocal ophthalmoscopy (cSLO) CX3CR1GFP/+ reporter mice, cellular resolution. Also, we describe quantify monthly cell density large subsets (200-300 cells per retina). We confirm use somal area as useful metric retina applying automated threshold-based morphometric analysis images. these image acquisition analyses strategies microgliosis early stages neurodegeneration mouse model glaucoma. approach should investigate contributions neuronal axonal decline disorders that affect optic nerve.