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Circadian oscillators in the mouse brain: molecular clock components in the neocortex and cerebellar cortex

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Abstract

The circadian timekeeper of the mammalian brain resides in the suprachiasmatic nucleus of the hypothalamus (SCN), and is characterized by rhythmic expression of a set of clock genes with specific 24-h daily profiles. An increasing amount of data suggests that additional circadian oscillators residing outside the SCN have the capacity to generate peripheral circadian rhythms. We have recently shown the presence of SCN-controlled oscillators in the neocortex and cerebellum of the rat. The function of these peripheral brain clocks is unknown, and elucidating this could involve mice with conditional cell-specific clock gene deletions. This prompted us to analyze the molecular clockwork of the mouse neocortex and cerebellum in detail. Here, by use of in situ hybridization and quantitative RT-PCR, we show that clock genes are expressed in all six layers of the neocortex and the Purkinje and granular cell layers of the cerebellar cortex of the mouse brain. Among these, Per1, Per2, Cry1, Arntl, and Nr1d1 exhibit circadian rhythms suggesting that local running circadian oscillators reside within neurons of the mouse neocortex and cerebellar cortex. The temporal expression profiles of clock genes are similar in the neocortex and cerebellum, but they are delayed by 5 h as compared to the SCN, suggestively reflecting a master–slave relationship between the SCN and extra-hypothalamic oscillators. Furthermore, ARNTL protein products are detectable in neurons of the mouse neocortex and cerebellum, as revealed by immunohistochemistry. These findings give reason to further pursue the physiological significance of circadian oscillators in the mouse neocortex and cerebellum.

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Abbreviations

Actb:

Actin beta

ANOVA:

Analysis of variance

Arntl:

Aryl hydrocarbon receptor nuclear translocator-like, also known as Bmal1

Clock:

Circadian locomotor output cycles kaput

Cry1:

Cryptochrome 1

CT:

Circadian time (animals kept in constant darkness for two days and sacrificed in darkness)

Dbp:

D site of albumin promoter binding protein

Gapdh:

Glyceraldehyde-3-phosphate dehydrogenase

GFAP:

Glial fibrillary acidic protein

NeuN:

Neuronal nuclear antigen

Nr1d1:

Nuclear receptor subfamily 1 group D member 1, also known as Rev-ErbAlpha

Per1:

Period circadian clock 1

Per2:

Period circadian clock 2

qRT-PCR:

Quantitative real-time RT-PCR

ZT:

Zeitgeber time (animals sacrificed during the light–dark cycle)

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Acknowledgments

This study was supported by the Lundbeck Foundation (R34-A3364 and R108-A10301 to M.F.R.; R67-A6494 to M.M.), the Danish Medical Research Council (271-09-0206 to M.F.R.) and the Simon Fougner Hartmann’s Family Foundation (to M.M.). We wish to thank Tine Thorup Mellergaard for expert technical assistance.

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  1. Department of Neuroscience and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Rigshospitalet 6102, Blegdamsvej 9, Copenhagen, DK-2100, Denmark

    Martin F. Rath, Louise Rovsing & Morten Møller

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  1. Martin F. Rath
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  2. Louise Rovsing
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  3. Morten Møller
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Correspondence to Martin F. Rath.

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Rath, M.F., Rovsing, L. & Møller, M. Circadian oscillators in the mouse brain: molecular clock components in the neocortex and cerebellar cortex. Cell Tissue Res 357, 743–755 (2014). https://doi.org/10.1007/s00441-014-1878-9

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