The retinal environment induces microglia-like properties in recruited myeloid cells

Background: Microglia are essential to the development of the CNS and its homeostasis. Our prior findings suggested a niche model to describe the behaviors of retinal microglia. Here, we ask whether new myeloid cells recruited to the retina are constrained to resemble endogenous microglia morphologically and functionally. Methods: Use of CD11c^DTR/GFP transgenic mouse allowed identification of two niches of retinal microglia distinguished by being GFP^lo or GFP^hi. We also used transgenic mice in which CX3CR1⁺ cells expressed YFP and were depletable following tamoxifen-induced expression of diphtheria toxin subunit A. We employed several ablation and injury stimulation protocols to examine the origin and fate of myeloid cells repopulating the retina. Analysis of retinal myeloid cells was done by microscopy, flow cytometry, and qRT-PCR. Results: We found that the origin of new GFP^hi and GFP^lo myeloid cells in the retina of CD11c^DTR/GFP mice, whether recruited or local, depended on the ablation and stimulation protocols. Regardless of origin, new GFP^lo and GFP^hi retinal myeloid cells were CD45^medCD11b⁺Ly6G^-Ly6C^loIba1⁺F4/80⁺, similar to endogenous microglia. Following tamoxifen-induced diphtheria toxin ablation, myeloid cell repopulation differed in the retina compared to the brain and optic nerve. Stimulation of replacement GFP^hi cells was substantially attenuated in repopulating retinas after tamoxifen-induced diphtheria toxin ablation compared to control or radiation-ablated mice. In radiation bone marrow chimeric mice, replacement GFP^hi myeloid cells from the circulation were slow to repopulate the retina unless stimulated by an optic nerve crush injury. However, once stimulated, recruited GFP^hi cells were found to concentrate on injured retinal ganglion cells and were morphologically similar to GFP^hi cells in non-ablated control CD11c^DTR/GFP mice. Conclusions: The results support the idea that GFP^hi cells in the CD11c^DTR/GFP mouse, whether recruited or from resident microglia, mark a unique niche of activated retinal myeloid cells. We conclude that the retinal environment has a potent influence on the function, morphology, and proliferative capacity of new myeloid cells regardless of their origin, compelling them to be equivalent to the endogenous microglia.