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ACCEPTED MANUSCRIPT However, due to the lack of understanding of the precise cellular signalling events during 670nm irradiation, there are still roadblocks in the translation of 670nm light therapy to the clinic (Hamblin, 2016). Previously we have demonstrated in the PD model that Müller cells, the principal macroglia of the retina, are the key source of the potent chemokine Ccl2 in retinal (Rutar et al., 2012; Rutar et al., 2011a), which is responsible for the recruitment and activation of MG/MΦ in retinas (Grigsby et al., 2014; Natoli et al., 2017b; Rutar et al., 2015). We postulated that the anti-inflammatory effect of 670nm may be due to a reduction of Müller cell-mediated inflammation during retinal degeneration. Our previous in vivo study suggested that pre-treatment with 670nm light mitigated photo-oxidative damage-induced structural changes in Müller cells (Albarracin and Valter, 2012). However, this model did not allow us to investigate the direct effects of 670nm light on the activation of Müller cells. In this study, by using a co-culture system where primary Müller cells are exposed to the environment of damaged photoreceptors, we can mimic gliotic changes that occur during photoreceptor degeneration in vivo, while allowing us to investigate the effects of PBM directly on the Müller cells. 2. Materials and Methods 2.1 Animals and light exposure All procedures were conducted in accordance with the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research and with ethics approval from the Australian National University Animal Experimentation Ethics Committee (Ethics ID: A2014/56). Albino Sprague-Dawley (SD) rats were born and raised in low light levels (5 lux) in a 12- hour light, 12-hour dark cycle. Food and water were available ad libitum. Twenty adult animals aged 100-120 postnatal (P) days were used for all experiments. Animals were separated into 4 experimental and control groups as follows: Control (n=5); Control + 670nm 5 ACCEPTED MANUSCRIPTPDF Image | Photobiomodulation with 670 nm light ameliorates retinal
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