Sphingosine-1-phosphate signaling through Müller glia regulates neuroprotection, accumulation of immune cells, and neuronal regeneration in the rodent retina.

The purpose of this study was to investigate how Sphingosine-1-phosphate (S1P) signaling regulates glial phenotype, neuroprotection, and reprogramming of Müller glia (MG) into neurogenic MG-derived progenitor cells (MGPCs) in the adult male and female mouse retina. We found that S1P-related genes were dynamically regulated following retinal damage. S1pr1 (S1P receptor 1) and Sphk1 (sphingosine kinase 1) are expressed at low levels by resting MG and are rapidly upregulated following acute damage. Overexpression of the neurogenic bHLH transcription factor Ascl1 in MG downregulates S1pr1, and inhibition of Sphk1 and S1pr1/3 enhances Ascl1-driven differentiation of bipolar-like cells. Treatments that activate S1pr1 or increase retinal levels of S1P initiate pro-inflammatory NFκB-signaling in MG, whereas treatments that inhibit S1pr1 or decreased levels of S1P suppress NFκB-signaling in MG. Conditional knock-out (cKO) of S1pr1 in MG, but not Sphk1, enhances the accumulation of immune cells in damaged retinas. cKO of S1pr1 promotes the survival of ganglion cells, whereas cKO of Sphk1 promotes the survival amacrine cells in damaged retinas. Consistent with these findings, pharmacological treatments that inhibit S1P receptors or inhibit Sphk1 had protective effects upon inner retinal neurons. We conclude that the S1P-signaling pathway is activated in MG after damage and this pathway acts secondarily to restrict the accumulation of immune cells, impairs neuron survival and suppresses the reprogramming of MG into neurogenic progenitors in the adult mouse retina.Significance Statement Understanding the mechanisms of retinal neuron survival and regeneration is fundamental for the development of therapeutic strategies of retinal repair. Here, we show that Sphingosine-1-phosphate signaling kick-starts the glial pro-inflammatory response, restricts immune cell recruitment, and exacerbates neuron cell death after acute retinal injury. Importantly, blocking sphingosine-1-phosphate activity enhances Ascl1-driven neurogenesis in the mouse retina, highlighting a potential therapeutic target for retinal regeneration.