A Robust Model System for Retinal Hypoxia: Live Imaging of Calcium Dynamics and Gene Expression Studies in Primary Human Mixed Retinal Culture.

摘要: The detailed mechanisms underlying oxidative stress that leads to neuroinflammation and neurodegeneration in retinal vascular conditions, including diabetic retinopathy, retinopathy of prematurity etc., remain largely unexplored mainly due a lack suitable disease models can simulate the inherent neuron-glia interactions human retina. Specifically, establishment mixed culture (MRC) containing both neuron glial cell types remains challenge different conditions required for their optimal growth differentiation. Here, we establish novel primary MRC model system neurons, astrocytes, Muller glia, microglia from donor retina be used study neuromodulatory effects cells under stress. characterization based on immunostaining with individual type-specific markers presence close vicinity each other further underscores utility studying cross talk. To best our knowledge, this is first instance an vitro obtained four major types. Next, induce hypoxic investigate if hypoxia activated neuroglia modulates altered gene expression inflammatory, apoptotic, angiogenic Ca2+ transients by live imaging. Further, performed k-means clustering responses identify modification pattern stressed condition. Finally, provide evidence transient correlates differential genes shown involved neuroinflammation, angiogenesis, as seen earlier lines animal retinopathy. features proposed included: increase activity, chemokine cytokine expression, percentage having higher amplitude frequency transients. Thus, experimental potentially serve ideal neuroinflammatory neurodegenerative changes identifying newer drug targets.