Abstract

As many as 10% of Americans suffer chronic sleep disturbances, but the genetic mechanisms that control sleep and wake states remain largely unknown. The long-range goal of the proposed studies is to identify secreted proteins that modulate sleep, wakefulness and arousal. Zebrafish will be used as a model system because it displays clear, measurable sleep and wake behaviors and is the only vertebrate system in which such a screen is feasible. The screen will involve the generation of several hundred stably transgenic zebrafish lines that express secreted proteins under an inducible promoter. The library will be made available through the zebrafish stock center and screened for modulators of larval sleep and wakefulness. In addition, the response to mechanical, chemical, thermal and visual cues will be tested. Candidate lines will be further analyzed in secondary screens for developmental and behavioral defects. The isolated peptides will form the foundation to dissect the molecular and neural circuits of sleep. The screen will also provide candidate pathways that might be affected in human sleep disorders and that could be modulated by drugs to treat sleep disorders.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL109525-05
Application #
8301000
Study Section
Special Emphasis Panel (ZRG1-CB-Z (57))
Program Officer
Laposky, Aaron D
Project Start
2008-06-01
Project End
2016-06-30
Budget Start
2012-07-01
Budget End
2013-06-30
Support Year
5
Fiscal Year
2012
Total Cost
$422,500
Indirect Cost
$172,500

  Institution

Name
Harvard University
Department
Microbiology/Immun/Virology
Type
Schools of Arts and Sciences
DUNS #
082359691
City
Cambridge
State
MA
Country
United States
Zip Code
02138

  Publications

Schoppik, David; Bianco, Isaac H; Prober, David A et al. (2017) Gaze-Stabilizing Central Vestibular Neurons Project Asymmetrically to Extraocular Motoneuron Pools. J Neurosci 37:11353-11365
Merkle, Florian T; Ghosh, Sulagna; Kamitaki, Nolan et al. (2017) Human pluripotent stem cells recurrently acquire and expand dominant negative P53 mutations. Nature 545:229-233
Escamilla, Christine Ochoa; Filonova, Irina; Walker, Angela K et al. (2017) Kctd13 deletion reduces synaptic transmission via increased RhoA. Nature 551:227-231
Santos, David P; Kiskinis, Evangelos; Eggan, Kevin et al. (2016) Comprehensive Protocols for CRISPR/Cas9-based Gene Editing in Human Pluripotent Stem Cells. Curr Protoc Stem Cell Biol 38:5B.6.1-5B.6.60
Thyme, Summer B; Akhmetova, Laila; Montague, Tessa G et al. (2016) Internal guide RNA interactions interfere with Cas9-mediated cleavage. Nat Commun 7:11750
Chiu, Cindy N; Rihel, Jason; Lee, Daniel A et al. (2016) A Zebrafish Genetic Screen Identifies Neuromedin U as a Regulator of Sleep/Wake States. Neuron 89:842-56
Thyme, Summer B; Schier, Alexander F (2016) Polq-Mediated End Joining Is Essential for Surviving DNA Double-Strand Breaks during Early Zebrafish Development. Cell Rep 15:707-714
Haesemeyer, Martin; Robson, Drew N; Li, Jennifer M et al. (2015) The structure and timescales of heat perception in larval zebrafish. Cell Syst 1:338-348
Merkle, Florian T; Maroof, Asif; Wataya, Takafumi et al. (2015) Generation of neuropeptidergic hypothalamic neurons from human pluripotent stem cells. Development 142:633-43
Merkle, Florian T; Neuhausser, Werner M; Santos, David et al. (2015) Efficient CRISPR-Cas9-mediated generation of knockin human pluripotent stem cells lacking undesired mutations at the targeted locus. Cell Rep 11:875-883

Showing the most recent 10 out of 25 publications