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ACCEPTED MANUSCRIPT Bringmann and Wiedemann, 2012). Mice deficient in GFAP exhibit less glial scars after retinal detachment (Nakazawa et al., 2007; Verardo et al., 2008), indicating a reduced activation of Müller cells. This may lead to the reduced expression of CCL2 and subsequent macrophage infiltration into retinas following damage (Nakazawa et al., 2007). 4.2 670nm light reduces Müller cell-mediated activation of MG/MΦ In the present study, we found the rapid upregulation of Ccl2 and Il-1β in the PD model in vivo, and by using a co-culture system of 661W photoreceptors and primary Müller cells in vitro, we have confirmed that the source of these factors are the Müller cells. Furthermore, using supernatant from activated Müller cells, we were able to initiate activation of MG/MΦ, confirming the direct link between Müller cell and macrophage activation. We have previously demonstrated that inhibition of Ccl2 and Il-1β reduces the infiltration of microglia and macrophages into the damaged outer retina in PD models (Natoli et al., 2017b; Rutar et al., 2012). The accumulation of MG/MΦ in the subretinal space is a well-established feature of retinal degenerations (Fernando et al., 2016; Knickelbein et al., 2015), which is associated with photoreceptor death (Natoli et al., 2017a; Zhao et al., 2015). The present study has demonstrated that treatment with 670nm light mitigates Ccl2 and Il-1β expression in the retina in vivo, and in primary Müller cells in vitro. Further, we found that 670nm light-treated Müller cells were able to mitigate MG/MΦ activation. This indicates that 670nm light targets the expression of pro-inflammatory cytokines and chemokines by Müller cells. Previous studies have found that 670nm light resulted in reduced activation and recruitment of MG/MΦ in retinal damage in vivo (Albarracin et al., 2011; Begum et al., 2013; Kokkinopoulos, 2013a; Kokkinopoulos et al., 2013b), but to our knowledge, this is the first report that demonstrated the direct effect of 670nm light on Müller cells and its downstream effect of reduced MG/MΦ activation. 18 ACCEPTED MANUSCRIPTPDF Image | Photobiomodulation with 670 nm light ameliorates retinal
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