Validation experiment designed to validate causal mechanisms targeting Pde6b in rd10 mouse organotypic retinal explants. Primary outcome: photoreceptor cell death reduction
Organotypic retinal explants from Pde6b(rd10) mice were treated with IGF-I to assess neuroprotective effects on photoreceptor cell death. The study evaluated cell death using TUNEL staining and ELISA of free nucleosomes, while monitoring microglial cell distribution using Cd11b and Iba1 immunolabeling. Microglia were depleted using clodronate-containing liposomes to determine their role in IGF-I-mediated neuroprotection. Results showed that IGF-I treatment reduced TUNEL-positive nuclei and increased microglial cells in the outer nuclear layer (ONL). Microglial depletion diminished IGF-I's neuroprotective effect but also moderately reduced baseline photoreceptor death.
Organotypic retinal explants treated with IGF-I, cell death assessed by TUNEL staining and ELISA of free nucleosomes, microglial visualization by Cd11b and Iba1 immunolabeling, microglial depletion with clodronate liposomes
IGF-I treatment would reduce photoreceptor cell death and modulate microglial response
Reduction in TUNEL-positive nuclei and free nucleosome levels
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