Desynchronizing the sleep­­–wake cycle from circadian timing to assess their separate contributions to physiology and behaviour and to estimate intrinsic circadian period

Wei Wang; Robin K. Yuan; Jude F. Mitchell; Kirsi-Marja Zitting; Melissa A. St. Hilaire; James K. Wyatt; Frank A. J. L. Scheer; Kenneth P. Wright; Emery N. Brown; Joseph M. Ronda; Elizabeth B. Klerman; Jeanne F. Duffy; Derk-Jan Dijk; Charles A. Czeisler

doi:10.1038/s41596-022-00746-y

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Desynchronizing the sleep–wake cycle from circadian timing to assess their separate contributions to physiology and behaviour and to estimate intrinsic circadian period

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Desynchronizing the sleep–wake cycle from circadian timing to assess their separate contributions to physiology and behaviour and to estimate intrinsic circadian period

Wei Wang, Robin K. Yuan, Jude F. Mitchell, Kirsi-Marja Zitting, Melissa A. St. Hilaire, James K. Wyatt, Frank A. J. L. Scheer, Kenneth P. Wright, Emery N. Brown, Joseph M. Ronda, …

Nature Protocols, Vol.18(2), pp.579-603

02/2023

DOI: https://doi.org/10.1038/s41596-022-00746-y

PMID: 36376588

Abstract

631/1647/2198

631/378/1385

692/308

Biological Techniques

Biomedical and Life Sciences

Computational Biology/Bioinformatics

General

Microarrays

Protocol

Behavioural methods

Circadian rhythms and sleep

Medical research

Analytical Chemistry

Life Sciences

Organic Chemistry

Circadian clocks drive cyclic variations in many aspects of physiology, but some daily variations are evoked by periodic changes in the environment or sleep–wake state and associated behaviors, such as changes in posture, light levels, fasting or eating, rest or activity and social interactions; thus, it is often important to quantify the relative contributions of these factors. Yet, circadian rhythms and these evoked effects cannot be separated under typical 24-h day conditions, because circadian phase and the length of time awake or asleep co-vary. Nathaniel Kleitman’s forced desynchrony (FD) protocol was designed to assess endogenous circadian rhythmicity and to separate circadian from evoked components of daily rhythms in multiple parameters. Under FD protocol conditions, light intensity is kept low to minimize its impact on the circadian pacemaker, and participants have sleep–wake state and associated behaviors scheduled to an imposed non-24-h cycle. The period of this imposed cycle, Τ , is chosen so that the circadian pacemaker cannot entrain to it and therefore continues to oscillate at its intrinsic period ( τ , ~24.15 h), ensuring circadian components are separated from evoked components of daily rhythms. Here we provide detailed instructions and troubleshooting techniques on how to design, implement and analyze the data from an FD protocol. We provide two procedures: one with general guidance for designing an FD study and another with more precise instructions for replicating one of our previous FD studies. We discuss estimating circadian parameters and quantifying the separate contributions of circadian rhythmicity and the sleep–wake cycle, including statistical analysis procedures and an R package for conducting the non-orthogonal spectral analysis method that enables an accurate estimation of period, amplitude and phase. This protocol describes how to design, implement and analyze the data from a forced desynchrony protocol to assess endogenous circadian rhythmicity and to separate circadian from evoked components of daily rhythms in physiology and behavior.

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Title: Desynchronizing the sleep–wake cycle from circadian timing to assess their separate contributions to physiology and behaviour and to estimate intrinsic circadian period
Creators: Wei Wang (Corresponding Author) - Brigham and Women's Hospital
Robin K. Yuan - Brigham and Women's Hospital
Jude F. Mitchell - University of Rochester
Kirsi-Marja Zitting - Brigham and Women's Hospital
Melissa A. St. Hilaire - Harvard Medical School
James K. Wyatt - Rush University Medical Center
Frank A. J. L. Scheer - Brigham and Women's Hospital
Kenneth P. Wright - University of Colorado Boulder
Emery N. Brown - Massachusetts General Hospital
Joseph M. Ronda - Brigham and Women's Hospital
Elizabeth B. Klerman - Brigham and Women's Hospital
Jeanne F. Duffy - Brigham and Women's Hospital
Derk-Jan Dijk - University of Surrey, School of Biosciences
Charles A. Czeisler - Brigham and Women's Hospital
Publication Details: Nature Protocols, Vol.18(2), pp.579-603
Publisher: Nature Publishing
Number of pages: 25
First online publication date: 14/11/2022
Publication Date: 02/2023
Date accepted for publication: 24/06/2022
Grant note: The FD studies and data described here were collected in the Environmental Scheduling Facility (ESF) and the Intensive Physiological Monitoring (IPM) Unit at the Brigham and Women’s Hospital. Both facilities were part of the BWH General Clinical Research Center (supported by National Institutes of Health (NIH) Grant M01 RR002635), and the IPM was part of the Harvard Clinical and Translational Science Center (supported by NIH Award UL1 RR025758 and financial contributions from the Brigham and Women’s Hospital and from Harvard University and its affiliated academic health care centers). The development of the FD protocol and NOSA program and carrying out the FD studies described here were supported by NIH Grants P01 AG09975, R01 HL080978, R01 HL52992, R21 AT02571, U01 AG12642 and T32 HL07901; by National Aeronautics and Space Administration Grants NAS9-19435 and NAG 5-3952 and NASA Cooperative Agreement NCC9-58 with the National Space Biomedical Research Institute; and by Air Force Office of Scientific Research Grants F49620-94-1-0398, F49620-95-1-0388, FA9550-06-0080 and F49620-00-1-0266.
Identifiers: 99668866502346; WOS:000885709500001
Copyright: © 2022, Springer Nature Limited. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
Academic Unit: School of Biosciences
Language: English
Resource Type: Journal article
Data Access Statement: All original datasets used in the preparation of this paper contain confidential or identifiable human data and are not publicly available due to human subject research regulations. De-identified sample datasets are available upon reasonable request and are subject to internal review and approval. Execution of a materials transfer agreement is required by our institution for transfer of data. SAS codes for data analyses and R codes for the NOSA program can be found at https://github.com/wwang4bwh/FD-NOSA.