Research

SCN-AVP release of mPer1/mPer2 double-mutant mice in vitro

Daan R van der Veen^1,3 , Ellis GA Mulder¹ , Henrik Oster² , Menno P Gerkema¹ and Roelof A Hut¹

¹  Department of Chronobiology, University of Groningen, P.O. Box 14, 9750 AA Haren, The Netherlands

²  Circadian Rhythms Group, Max Planck Institute of Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen, Germany

³  University of Surrey, Faculty of Health and Medical Sciences, Guildford, Surrey GU2 7XH, UK

author email corresponding author email

Journal of Circadian Rhythms 2008, 6:5doi:10.1186/1740-3391-6-5

Published:	20 March 2008

Abstract

Background

Circadian organisation of behavioural and physiological rhythms in mammals is largely driven by the clock in the suprachiasmatic nuclei (SCN) of the hypothalamus. In this clock, a molecular transcriptional repression and activation mechanism generates near 24 hour rhythms. One of the outputs of the molecular clock in specific SCN neurons is arginine-vasopressin (AVP), which is responsive to transcriptional activation by clock gene products. As negative regulators, the protein products of the period genes are thought to repress transcriptional activity of the positive limb after heterodimerisation with CRYPTOCHROME. When both the Per1 and Per2 genes are dysfunctional by targeted deletion of the PAS heterodimer binding domain, mice lose circadian organization of behaviour upon release into constant environmental conditions. To which degree the period genes are involved in the control of AVP output is unknown.

Methods

Using an in vitro slice culture setup, SCN-AVP release of cultures made of 10 wildtype and 9 Per1/2 double-mutant mice was assayed. Mice were sacrificed in either the early light phase of the light-dark cycle, or in the early subjective day on the first day of constant dark.

Results

Here we report that in arrhythmic homozygous Per1/2 double-mutant mice there is still a diurnal peak in in vitro AVP release from the SCN similar to that of wildtypes but distinctively different from the release pattern from the paraventricular nucleus. Such a modulation of AVP release is unexpected in mice where the circadian clockwork is thought to be disrupted.

Conclusion

Our results suggest that the circadian clock in these animals, although deficient in (most) behavioural and molecular rhythms, may still be (partially) functional, possibly as an hourglass mechanism. The level of perturbation of the clock in Per1/2 double mutants may therefore be less than was originally thought.