Circadian oscillator function in embryonic retina and retinal explant cultures

Retinal circadian oscillators regulate many aspects of retinal function. Investigations of these oscillators and the biochemical cascades that entrain them would be greatly facilitated if experimental paradigms could be identified that permit long-term monitoring of retinal circadian oscillator function in vitro. The purpose of this study was to determine if chicken retinas maintained in explant culture conditions could serve in this capacity. Retinal circadian oscillator function was studied by monitoring iodopsin transcription under cyclic light, constant dark, and following reversal of the light cycle. Rhythms observed in the explant cultures were compared to those observed in retinas of embryos (in ovo) and post-hatch chickens. Robust iodopsin transcript rhythms were observed for up to 9 days in explant cultures maintained under cyclic light. These rhythms persisted for 48 h in constant darkness and the time course for re-entrainment of the rhythm to a reversed light/dark cycle was similar to that observed in post-hatch chicken retinas. These results show that circadian oscillators located within the retina play a key role in the regulation of iodopsin transcription in retinal explant cultures and in retinas of post-hatch chickens. Interestingly, our data show that iodopsin transcription in retinas of intact embryos is primarily, if not entirely, driven by light. These results show that the circadian oscillators driving iodopsin transcription in embryonic retinal explant cultures exhibit functional characteristics similar to those found in post-hatch chicken retina, supporting use of this paradigm in further studies of entrainment of these oscillators in retina.