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Live imaging of altered period1 expression in the suprachiasmatic nuclei of Vipr2⁻/⁻ mice
Vasoactive intestinal polypeptide and its receptor, VPAC₂, play important roles in the functioning of the brain's circadian clock in the suprachiasmatic nuclei (SCN). Mice lacking VPAC₂ receptors (Vipr2⁻/⁻) show altered circadian rhythms in locomotor behavior, neuronal firing rate, and clock ge...
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Published in: | Journal of neurochemistry 2008-08, Vol.106 (4), p.1646-1657 |
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Main Authors: | , , , , , |
Format: | Article |
Language: | English |
Subjects: | |
Online Access: | Get full text |
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Summary: | Vasoactive intestinal polypeptide and its receptor, VPAC₂, play important roles in the functioning of the brain's circadian clock in the suprachiasmatic nuclei (SCN). Mice lacking VPAC₂ receptors (Vipr2⁻/⁻) show altered circadian rhythms in locomotor behavior, neuronal firing rate, and clock gene expression, however, the nature of molecular oscillations in individual cells is unclear. Here, we used real-time confocal imaging of a destabilized green fluorescent protein (GFP) reporter to track the expression of the core clock gene Per1 in live SCN-containing brain slices from wild-type (WT) and Vipr2⁻/⁻ mice. Rhythms in Per1-driven GFP were detected in WT and Vipr2⁻/⁻ cells, though a significantly lower number and proportion of cells in Vipr2⁻/⁻ slices expressed detectable rhythms. Further, Vipr2⁻/⁻ cells expressed significantly lower amplitude oscillations than WT cells. Within each slice, the phases of WT cells were synchronized whereas cells in Vipr2⁻/⁻ slices were poorly synchronized. Most GFP-expressing cells, from both genotypes, expressed neither vasopressin nor vasoactive intestinal polypeptide. Pharmacological blockade of VPAC₂ receptors in WT SCN slices partially mimicked the Vipr2⁻/⁻ phenotype. These data demonstrate that intercellular communication via the VPAC₂ receptor is important for SCN neurons to sustain robust, synchronous oscillations in clock gene expression. |
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ISSN: | 0022-3042 1471-4159 |
DOI: | 10.1111/j.1471-4159.2008.05520.x |