Large-scale Analysis of Sleep in Zebrafish

Overview

Journal Bio Protoc

Specialty Biology

Date 2022 Mar 14

PMID 35284597

Authors

Daniel A Lee

Grigorios Oikonomou

David A Prober

Affiliations

Soon will be listed here.

Abstract

Over the past decade, zebrafish have emerged as a powerful model for the study of vertebrate sleep and wake behaviors. Experimental evidence has demonstrated behavioral, anatomical, genetic, and pharmacological conservation of sleep between zebrafish and mammals, suggesting that discoveries in zebrafish can inform our understanding of mammalian sleep. Here, we describe a protocol for performing sleep behavioral experiments in larval zebrafish, using a high-throughput video tracking system. We explain how to set up a sleep behavioral experiment and provide guidelines on how to analyze the data. Using this protocol, a typical experiment can be completed in less than five days, and this method provides a scalable platform to perform genetic and pharmacological screens in a simple and cost-effective vertebrate model. By combining high-throughput behavioral assays with several advantageous features of zebrafish, this model system provides new opportunities to make discoveries that clarify the genetic and neurological mechanisms that regulate sleep.

Citing Articles

Behavioural pharmacology predicts disrupted signalling pathways and candidate therapeutics from zebrafish mutants of Alzheimer's disease risk genes.

Kroll F, Donnelly J, Ozcan G, Mackay E, Rihel J Elife. 2025; 13.

PMID: 39960847 PMC: 11832171. DOI: 10.7554/eLife.96839.

Measuring Sleep and Activity Patterns in Adult Zebrafish.

Doldur-Balli F, Zimmerman A, Seiler C, Veatch O, Pack A Bio Protoc. 2024; 14(12):e5014.

PMID: 38948256 PMC: 11211082. DOI: 10.21769/BioProtoc.5014.

Screening effects of HCN channel blockers on sleep/wake behavior in zebrafish.

Doldur-Balli F, Smieszek S, Keenan B, Zimmerman A, Veatch O, Polymeropoulos C Front Neurosci. 2024; 18:1375484.

PMID: 38567282 PMC: 10986788. DOI: 10.3389/fnins.2024.1375484.

Pleiotropic effects of a high confidence Autism Spectrum Disorder gene, , on zebrafish sleep.

Doldur-Balli F, Zimmerman A, Keenan B, Shetty Z, Grant S, Seiler C Neurobiol Sleep Circadian Rhythms. 2023; 14:100096.

PMID: 37287661 PMC: 10241967. DOI: 10.1016/j.nbscr.2023.100096.

Identification of nicotine-seeking and avoiding larval zebrafish using a new three-choice behavioral assay.

Schneider H, Pearson A, Harris D, Krause S, Tucker A, Gardner K Front Mol Neurosci. 2023; 16:1112927.

PMID: 37063370 PMC: 10098024. DOI: 10.3389/fnmol.2023.1112927.

References

Cirelli C, Bushey D, Hill S, Huber R, Kreber R, Ganetzky B . Reduced sleep in Drosophila Shaker mutants. Nature. 2005; 434(7037):1087-92. DOI: 10.1038/nature03486. View

Hendricks J, Finn S, Panckeri K, Chavkin J, Williams J, Sehgal A . Rest in Drosophila is a sleep-like state. Neuron. 2000; 25(1):129-38. DOI: 10.1016/s0896-6273(00)80877-6. View

Ahrens M, Li J, Orger M, Robson D, Schier A, Engert F . Brain-wide neuronal dynamics during motor adaptation in zebrafish. Nature. 2012; 485(7399):471-7. PMC: 3618960. DOI: 10.1038/nature11057. View

Singh C, Rihel J, Prober D . Neuropeptide Y Regulates Sleep by Modulating Noradrenergic Signaling. Curr Biol. 2017; 27(24):3796-3811.e5. PMC: 5736401. DOI: 10.1016/j.cub.2017.11.018. View

Lee D, Andreev A, Truong T, Chen A, Hill A, Oikonomou G . Genetic and neuronal regulation of sleep by neuropeptide VF. Elife. 2017; 6. PMC: 5705210. DOI: 10.7554/eLife.25727. View

Reichert S, Arocas O, Rihel J . The Neuropeptide Galanin Is Required for Homeostatic Rebound Sleep following Increased Neuronal Activity. Neuron. 2019; 104(2):370-384.e5. DOI: 10.1016/j.neuron.2019.08.010. View

Chiu C, Rihel J, Lee D, Singh C, Mosser E, Chen S . A Zebrafish Genetic Screen Identifies Neuromedin U as a Regulator of Sleep/Wake States. Neuron. 2016; 89(4):842-56. PMC: 4851465. DOI: 10.1016/j.neuron.2016.01.007. View

Gandhi A, Mosser E, Oikonomou G, Prober D . Melatonin is required for the circadian regulation of sleep. Neuron. 2015; 85(6):1193-9. PMC: 4851458. DOI: 10.1016/j.neuron.2015.02.016. View

Elbaz I, Yelin-Bekerman L, Nicenboim J, Vatine G, Appelbaum L . Genetic ablation of hypocretin neurons alters behavioral state transitions in zebrafish. J Neurosci. 2012; 32(37):12961-72. PMC: 6703801. DOI: 10.1523/JNEUROSCI.1284-12.2012. View

10.

Jeong J, Kwon H, Ahn J, Kang D, Kwon S, Park J . Functional and developmental analysis of the blood-brain barrier in zebrafish. Brain Res Bull. 2008; 75(5):619-28. DOI: 10.1016/j.brainresbull.2007.10.043. View

11.

Yelin-Bekerman L, Elbaz I, Diber A, Dahary D, Gibbs-Bar L, Alon S . Hypocretin neuron-specific transcriptome profiling identifies the sleep modulator Kcnh4a. Elife. 2015; 4:e08638. PMC: 4718730. DOI: 10.7554/eLife.08638. View

12.

Chen S, Reichert S, Singh C, Oikonomou G, Rihel J, Prober D . Light-Dependent Regulation of Sleep and Wake States by Prokineticin 2 in Zebrafish. Neuron. 2017; 95(1):153-168.e6. PMC: 5653285. DOI: 10.1016/j.neuron.2017.06.001. View

13.

Prober D, Rihel J, Onah A, Sung R, Schier A . Hypocretin/orexin overexpression induces an insomnia-like phenotype in zebrafish. J Neurosci. 2006; 26(51):13400-10. PMC: 6675014. DOI: 10.1523/JNEUROSCI.4332-06.2006. View

14.

Shaw P, Cirelli C, Greenspan R, Tononi G . Correlates of sleep and waking in Drosophila melanogaster. Science. 2000; 287(5459):1834-7. DOI: 10.1126/science.287.5459.1834. View

15.

Ozcan G, Lim S, Leighton P, Allison W, Rihel J . Sleep is bi-directionally modified by amyloid beta oligomers. Elife. 2020; 9. PMC: 7360368. DOI: 10.7554/eLife.53995. View

16.

Fisher S, Godinho S, Pothecary C, Hankins M, Foster R, Peirson S . Rapid assessment of sleep-wake behavior in mice. J Biol Rhythms. 2012; 27(1):48-58. PMC: 4650254. DOI: 10.1177/0748730411431550. View

17.

Lee D, Liu J, Hong Y, Lane J, Hill A, Hou S . Evolutionarily conserved regulation of sleep by epidermal growth factor receptor signaling. Sci Adv. 2019; 5(11):eaax4249. PMC: 6853770. DOI: 10.1126/sciadv.aax4249. View

18.

Churgin M, Jung S, Yu C, Chen X, Raizen D, Fang-Yen C . Longitudinal imaging of in a microfabricated device reveals variation in behavioral decline during aging. Elife. 2017; 6. PMC: 5484621. DOI: 10.7554/eLife.26652. View

19.

Koh K, Joiner W, Wu M, Yue Z, Smith C, Sehgal A . Identification of SLEEPLESS, a sleep-promoting factor. Science. 2008; 321(5887):372-6. PMC: 2771549. DOI: 10.1126/science.1155942. View

20.

Renier C, Faraco J, Bourgin P, Motley T, Bonaventure P, Rosa F . Genomic and functional conservation of sedative-hypnotic targets in the zebrafish. Pharmacogenet Genomics. 2007; 17(4):237-53. DOI: 10.1097/FPC.0b013e3280119d62. View