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Fig. 3 | Acta Neuropathologica Communications

Fig. 3

From: Optic nerve as a source of activated retinal microglia post-injury

Fig. 3

Fate mapping showed that the retinal GFPhi myeloid cells post-ONC were derived from microglia. a Tam induction of the RFP reporter in CX3CR1YFP-CreER:R26RFP mice labeled circulating CD11b+CD45hi mononuclear cells, which substantially declined by 47 days. b Fundus photography showed that the retinal microglia were well labeled and expressed RFP at 70 days. c Flow cytometry showed that the microglia were induced to stably express RFP. (Left panels) An ONC-injured retina (ipsilateral) in mice lacking the CD11c-GFP transgene responded 7 days later with an increased number of microglia (YFPhiRFPhi) relative to the control (contralateral) retina. (Right panels) CX3CR1YFP-CreER:R26RFP:CD11cGFP mice upregulated GFP on the YFPhiRFPhi microglia in the ipsilateral ONC retinas, accounting for a substantial portion of the increase in retinal myeloid cells. d Quantitation of the flow cytometry from c showing averaged results of groups of 3 to 6 mice. All GFPhi cells were also RFPhi and YFPhi. Abbreviations: GFP−/− = CX3CR1YFP-CreER:R26RFP mice. GFP+/− = CX3CR1YFP-CreER:R26RFP:CD11cGFP mice. Flow cytometry on retina included gating for viability, doublet exclusion and FSC/SSC prior to gating on CD45medCD11b+Ly6G for expression of RFP, YFP and GFP. e Flatmount focused on the NFL of a retina post-ONC in a Tam-induced CX3CR1YFP-CreER:R26RFP:CD11cGFP mouse showed that the GFPhi cells were RFPhi

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