Dual-Color Single-Cell Imaging of the Suprachiasmatic Nucleus Reveals a Circadian Role in Network Synchrony

Yongli Shan; John H Abel; Yan Li; Mariko Izumo; Kimberly H Cox; Byeongha Jeong; Seung-Hee Yoo; David P Olson; Francis J Doyle 3rd; Joseph S Takahashi

doi:10.1016/j.neuron.2020.07.012

Dual-Color Single-Cell Imaging of the Suprachiasmatic Nucleus Reveals a Circadian Role in Network Synchrony

Neuron. 2020 Oct 14;108(1):164-179.e7. doi: 10.1016/j.neuron.2020.07.012. Epub 2020 Aug 7.

Authors

Affiliations

¹ Department of Neuroscience, Peter O'Donnell Jr. Brain Institute, University of Texas Southwestern Medical Center, Dallas, TX 75390-9111, USA.
² Department of Anesthesiology, Critical Care and Pain Medicine, Massachusetts General Hospital, Boston, MA 02114, USA; Department of Systems Biology, Harvard Medical School, Boston, MA 02115, USA; Picower Institute for Learning and Memory, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
³ Department of Pediatrics, Department of Molecular & Integrative Physiology, University of Michigan, Ann Arbor, MI 48109, USA.
⁴ Harvard John A. Paulson School of Engineering and Applied Sciences, Harvard University, Cambridge, MA 02138, USA; Division of Sleep Medicine, Harvard Medical School, Boston, MA 02115, USA.
⁵ Department of Neuroscience, Peter O'Donnell Jr. Brain Institute, University of Texas Southwestern Medical Center, Dallas, TX 75390-9111, USA; Howard Hughes Medical Institute, University of Texas Southwestern Medical Center, Dallas, TX 75390-9111, USA. Electronic address: joseph.takahashi@utsouthwestern.edu.

Abstract

The suprachiasmatic nucleus (SCN) acts as a master pacemaker driving circadian behavior and physiology. Although the SCN is small, it is composed of many cell types, making it difficult to study the roles of particular cells. Here we develop bioluminescent circadian reporter mice that are Cre dependent, allowing the circadian properties of genetically defined populations of cells to be studied in real time. Using a Color-Switch PER2::LUCIFERASE reporter that switches from red PER2::LUCIFERASE to green PER2::LUCIFERASE upon Cre recombination, we assess circadian rhythms in two of the major classes of peptidergic neurons in the SCN: AVP (arginine vasopressin) and VIP (vasoactive intestinal polypeptide). Surprisingly, we find that circadian function in AVP neurons, not VIP neurons, is essential for autonomous network synchrony of the SCN and stability of circadian rhythmicity.

Keywords: Per2 gene; SCN; bioluminescence imaging; circadian rhythms; luciferase; neuronal coupling; neuronal network; suprachiasmatic nucleus.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Video-Audio Media

MeSH terms

ARNTL Transcription Factors / genetics
Animals
Arginine Vasopressin / metabolism*
Circadian Rhythm*
Gene Knockout Techniques
Luciferases
Mice
Mice, Transgenic
Nerve Net / metabolism*
Period Circadian Proteins / genetics*
Period Circadian Proteins / metabolism
Single-Cell Analysis
Suprachiasmatic Nucleus / metabolism
Suprachiasmatic Nucleus Neurons / metabolism*
Time-Lapse Imaging
Vasoactive Intestinal Peptide / metabolism*

Substances

ARNTL Transcription Factors
Bmal1 protein, mouse
Per2 protein, mouse
Period Circadian Proteins
Arginine Vasopressin
Vasoactive Intestinal Peptide
Luciferases

Abstract

Publication types

MeSH terms

Substances

Grants and funding