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Assessment of Circadian and Light‐Entrainable Parameters in Mice Using Wheel‐Running Activity

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  • Abstract
  • Table of Contents
  • Materials
  • Figures
  • Literature Cited

Abstract

 

In most organisms, physiological variables are regulated by an internal clock. This endogenous circadian (?24?hr) clock enables organisms to anticipate daily environmental changes and modify behavioral and physiological functions appropriately. Processes regulated by the circadian clock include sleep?wake and locomotor activity, core body temperature, metabolism, water/food intake, and available hormone levels. At the core of the mammalian circadian system are molecular oscillations within the hypothalamic suprachiasmatic nucleus. These oscillations are modifiable by signals from the environment (so called zeitgebers or time?givers) and, once integrated within the suprachiasmatic nucleus, are conveyed to remote neural circuits where output rhythms are regulated. Disrupting any of a number of neural processes can affect how rhythms are generated and relayed to the periphery and disturbances in circadian/entrainment parameters are associated with numerous human conditions. These non?invasive protocols can be used to determine whether circadian/entrainment parameters are affected in mouse mutants or treatment groups. Curr. Protoc. Mouse Biol. 1:369?381 © 2011 by John Wiley & Sons, Inc.

Keywords: circadian; light entrainment; period; phase; amplitude; constant conditions

     
 
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Table of Contents

  • Introduction
  • Strategic Planning
  • Basic Protocol 1: Circadian Activity in Light/Dark Cycles and Constant Conditions
  • Support Protocol 1: Masking and Phase Shifting
  • Support Protocol 2: T‐Cycles and Reentrainment
  • Commentary
  • Literature Cited
  • Figures
     
 
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Materials

Basic Protocol 1: Circadian Activity in Light/Dark Cycles and Constant Conditions

  Materials
  • Mice (unless chambers being used are equipped with individually ventilated cages, IVC, all animals should be same sex; young adult mice between 8 and 20 weeks; cohorts of at least ten mice per genotype or treatment group)
  • Cages with running wheels, bedding, but no other environmental enrichment (running wheels equipped with system to quantify number of revolutions, e.g., available from http://www.coulbourn.com; http://www.panlab.com; http://www.techniplastuk.com; http://www.tse‐systems.com; http://www.lafayetteinstrument.com) or alternatively, use microswitches or magnets to custom make activity monitoring running wheels for any standard home cage (Jud et al., ; Siepka and Takahashi, ), individual microswitches are connected to a data‐collection computer via cabling connected to data acquisition boards
  • Circadian light‐tight chambers (see Strategic Planning; light set at ∼150 lux and estimated using a lux meter; air flow, temperature, and humidity maintained according to institutional recommendations)
  • Dedicated room for housing circadian monitoring system (a set of double doors useful to prevent light from accidentally entering circadian chambers during cage checking; cover all potential light sources within the room, e.g., power monitor lights, with light‐proof tape; cover overhead lighting with light filters, e.g., Kodak no. 11, to check mice daily in darkness without using infra‐red goggles)
  • Data collection computer and software to record wheel running activity (e.g., ClockLab, http://www.actimetrics.com/ClockLab/; VitalView, http://www.minimitter.com/vitalview_software.cfm; The Chronobiology Kit, http://www.query.com/chronokit/; or Med Associates SOF‐860, http://www.med‐associates.com/activity/wireless.htm)
  • Data collection hardware (National Instruments, cat. no. PCI‐6023E)
  • Data analysis computer (e.g., for ClockLab, an additional license for MATLAB software, The Mathworks, is required for data analysis; upload of data to the data analysis computer is automated to occur every 2 hr)
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Figures

  •   Figure Figure 1. Typical set‐up for recording wheel‐running activity in mice showing light‐controlled circadian chambers and wheel running cages. (A ) A full circadian chamber, open to reveal wheel running cages. (B ) Close up of wheel running cages within the circadian chamber. (C ) Singly housed mouse within a wheel running cage.
    View Image
  •   Figure Figure 2. Actograms. (A ) Standard actogram, showing wheel running activity over 2 days in a 12:12 hr light/dark cycle followed by 2 days in constant darkness. Each day's data is presented beneath that of the previous day. Each horizontal line represents 24 hr of activity and activity is represented by vertical bars. Actograms are shaded where lights are off. (B ) Double‐plotted actogram of the same data shown in A. Each horizontal line represents 48 hr of activity and each day's data is presented both beneath and to the right of the previous day.
    View Image
  •   Figure Figure 3. Data generated using the . (A ) Actogram showing mouse wheel running data using the conditions outlined in the . Actograms are shaded where lights are off. Note the activity relative to lights‐off in the first part of the protocol, the period shortening in constant darkness and period lengthening in constant light. (B ) Graph showing average wheel running counts in the different lighting conditions of the . (C ) Actogram showing splitting in the constant light phase of the screen. Actograms are shaded where lights are off.
    View Image
  •   Figure Figure 4. Data generated from . (A ) Actogram showing responses to light and dark pulses. Actograms are shaded where lights are off. Note the acute suppression of wheel‐running activity by light (masking). Light pulses can also result in sustained effects on the phase of activity. In this example, the mouse shows a delayed phase of activity for 6 days after the light pulse. (B ) Actogram showing the phase delay in the onset of activity in constant darkness following a 15‐min light pulse applied at CT16 (light pulse is indicated by an * on the actogram). (C ) Actogram showing phase advance in the onset of activity in constant darkness following a 15‐min light pulse applied at CT23 (light pulse is indicated by an * on the actogram). (D ) Representation of a phase response curve. Note the time window during subjective day (CT02‐CT12) where activity phase is not affected by light pulses.
    View Image
  •   Figure Figure 5. Data generated from . (A ) Actogram showing activity during a 12:12 followed by an 11:11 hr light/dark T‐cycle. Actogram is double‐plotted on a 24‐hr cycle. Actograms are shaded where lights are off. (B ) Actogram plotted using the same data as for A above, but the actogram is double plotted on a 22‐hr cycle. Actograms are shaded where lights are off. (C ) Actogram showing re‐entrainment following 6 hr advance and delay shifts in the light/dark cycle. Actograms are shaded where lights are off.
    View Image

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Literature Cited

Literature Cited
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