Daily rhythms of locomotor activity and transcript levels of non-visual opsins in the brain of the blind Mexican cavefish (Astyanax mexicanus)

Most organisms possess endogenous circadian clocks that synchronise their physiology and behaviour with environmental cycles, with the light-dark (LD) cycle being the most potent synchronising signal. Consequently, it can be hypothesised that animals that have evolved in the dark, as in caves or deep sea, may no longer possess a functional light-entrained biological clock. In this research, the blind cavefish Astyanax mexicanus was selected as a model organism to investigate the potential effects of daily light conditions on the circadian timekeeping mechanisms. First, we focused on describing behavioural photic entrainment and the presence of a circadian endogenous rhythmicity by recording locomotor activity rhythms under different lighting regimes: LD 12:12, after a 6-h shift of LD, constant darkness (DD), and constant dim light (LLdim). Secondly, we aimed at characterising the mechanisms of photodetection by analysing the daily rhythms of expression of selected non-visual extraocular opsins (exo-rhod, opn3, rgra, rgrb, tmt1a and tmt1b) in the brain of this blind species using real-time quantitative PCR. Our results revealed that blind Mexican cavefish activity rhythms were entrained to the LD cycle, with a diurnal activity pattern that persisted in a circadian fashion under constant lighting conditions. Additionally, statistically significant daily variations and/or rhythms were observed in three out of the six non-visual opsin genes analysed (opn3, rgra and tmt1b), all of them displaying nocturnal acrophases. These findings suggest that daily rhythms in extraretinal non-visual opsins may be transducing daily photic cycles and contributing to the entrainment of locomotor activity and other light-synchronised rhythms in blind cavefish species.