Wavelength-specific artificial light disrupts molecular clock in avian species: A power-calibrated statistical approach.

Nighttime lighting is an increasingly important anthropogenic environmental stress on plants and animals. Exposure to unnatural lighting environments may disrupt the circadian rhythm of organisms. However, the sample size of relevant studies, e.g. disruption of the molecular circadian clock by light pollution, was small (<10), which led to low statistical power and difficulties in replicating prior results. Here, we developed a power-calibrated statistical approach to overcome these weaknesses. The results showed that the effect size of 2.48 in clock genes expression induced by artificial light would ensure the reproducibility of the results as high as 80%. Long-wavelength light (560-660 nm) entrained expressions of the positive core clock genes (e.g. cClock) and negative core clock genes (e.g. cCry1, cPer2) in robust circadian rhythmicity, whereas those clock genes were arrhythmic in short-wavelength light (380-480 nm). Further, we found artificial light could entrain the transcriptional-translational feedback loop of the molecular clock in a wavelength-dependent manner. The expression of the positive core clock genes (cBmal1, cBmal2 and cClock), cAanat gene and melatonin were the highest in short-wavelength light and lowest in long-wavelength light. For the negative regulators of the molecular clock (cCry1, cCry2, cPer2 and cPer3), the expression of which was the highest in long-wavelength light and lowest in short-wavelength light. Our statistical approach opens new opportunities to understand and strengthen conclusions, comparing with the studies with small sample sizes. We also provide comprehensive insight into the effect of wavelength-specific artificial light on the circadian rhythm of the molecular clock in avian species. Especially, the global lighting is shifting from "yellow" sodium lamps, which is more like the long-wavelength light, toward short-wavelength light (blue light)-enriched "white" light-emitting diodes (LEDs).