Circadian regulation of the PERIOD 2::LUCIFERASE bioluminescence rhythm in the mouse retinal pigment epithelium-choroid.

Dec 15, 2010Molecular vision

Daily rhythms of a clock gene’s light signal in the mouse eye’s support layer

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Abstract

Per2 mRNA levels in the RPE-choroid show a clear circadian rhythm in vivo.

A circadian rhythm in bioluminescence was observed in cultured RPE-choroid explants from PER2::LUC knockin mice.
Light exposure during subjective night did not shift the phase of the circadian rhythm in bioluminescence.
Removal of the suprachiasmatic nuclei did not impact the circadian rhythm of bioluminescence in the RPE-choroid.
These findings suggest that the regulation of the circadian rhythm in the RPE-choroid is independent of traditional central circadian control.

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PURPOSE: The retinal pigment epithelium (RPE) plays an important role in the maintenance of the health and function of photoreceptors. Previous studies have shown that the RPE is also involved in the regulation of disc shedding, a process that is vital for photoreceptor health. This process has been shown to be under circadian control, although the mechanisms that control it are poorly understood. The aim of the present study was to investigate Period 2 (Per2) mRNA levels in the mouse RPE in vivo, and to determine whether the cultured RPE-choroid from PERIOD 2::LUCIFERASE (PER2::LUC) knockin mice expresses a circadian rhythm in bioluminescence.

METHODS: Per2 mRNA levels were measured using real-time quantitative RT-PCR, and bioluminescence was measured in PER2::LUC knockin mice using a Lumicycle®.

RESULTS: Per2 mRNA levels in the RPE-choroid show a clear circadian rhythm in vivo. A circadian rhythm in PER2::LUC bioluminescence was recorded from cultured RPE-choroid explants. Light exposure during the subjective night did not cause a circadian rhythm phase-shift of PER2::LUC bioluminescence. Finally, removal of the suprachiasmatic nuclei of the hypothalamus did not affect the bioluminescence circadian rhythm in the RPE-choroid.

CONCLUSIONS: Our results demonstrate that the RPE-choroid contains a circadian clock, and the regulation of this circadian rhythm resides within the eye. These new data indicate that it may be useful to design studies with the aim of elucidating the molecular mechanisms responsible for the regulation of the rhythmic event in the RPE.

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Circadian regulation of the PERIOD 2::LUCIFERASE bioluminescence rhythm in the mouse retinal pigment epithelium-choroid.

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