# Project 4: Whole-brain and body characterization of sleep disturbances and interventions in Fmr1, Shank3 and Cntnap2 knockout zebrafish

> **NIH NIH P50** · STANFORD UNIVERSITY · 2023 · $396,570

## Abstract

Project 4: Project Summary/Abstract
Sleep is critical for proper synaptic connections and brain development. Our group previously established that
sleep disruptions in zebrafish, like in other species, prevent normal structural synapse plasticity. Conversely,
proper sleep and melatonin hypnotic/circadian treatment can improve these synaptic defects. Animal models
of ASD, like ASD patients, suffer from sleep disruptions during development and display synaptic and
behavioral deficiencies. Here, we hypothesize that sleep disruptions during development are causal
and/or aggravating factors of ASD synaptic and behavioral defects, and that sleep interventions could
alleviate these issues. While human (Projects 1 & 2) and mouse (Project 3) approaches permit exquisite
studies of social interactions, repetitive behaviors, and associated cortical synaptic defects, zebrafish is a
transparent vertebrate popular in developmental biology allowing whole brain and body investigation.
Importantly, ASD risk genes like Fmr1, Shank3, and Cntnap2 are pan-neuronal, and their loss likely impacts
the entire central nervous system during sleep. We have recently developed fluorescence-based
polysomnography (fPSG) in zebrafish, a novel, non-invasive method allowing whole-brain and whole-body
imaging with single cell resolution during sleep. Using fPSG, we have shown that zebrafish have sleep brain
dynamics analogous to mammals, including a state we coined slow bursting sleep (SBS) which shares many
commonalities with Non-REM slow wave sleep (SWS). Our preliminary data indicates that SBS is fragmented
in developing Fmr1 zebrafish mutants. Further, studies from other groups have shown that based on actimetry,
sleep/wake pattern is also disrupted in zebrafish cntnapt2ab and shank3ab mutants. However, their brain
activity during sleep has not yet been investigated. Thus, in Aim 1, we will apply fPSG to these three genotypes
(fmr1, shank3ab, and cntnap2ab mutants) and controls to fully characterize their sleep neural and muscular
dynamics during development. Next, we will apply the same pharmacological interventions (H1R antihistamine,
GABAA agonist, and hypocretin/orexin receptors antagonist) used in human (Project 2) and mouse (Project
3), to improve sleep onset latency and sleep/SBS consolidation in these ASD risk gene mutants. Then, in Aim
2, we will investigate the respective beneficial effects of these NREM/SWS/SBS-sleep interventions on
structural synapse density using longitudinal imaging of telencephalic, hypothalamic and spinal cord circuits
expressing synaptic proteins fused to fluorescent markers such as PSD95-eGFP, Synaptophysin-eGFP or
Gephyrin-eGFP. In parallel, treated fish will be assessed for improvement in repetitive and social behaviors
like in mouse (Project 3) and human (Project 2). Complementing the latter, the transparency of the zebrafish
model will reveal how sleep dynamics are disrupted throughout the entire brain and how sleep interventions
can also ...

## Key facts

- **NIH application ID:** 10698080
- **Project number:** 5P50HD109861-02
- **Recipient organization:** STANFORD UNIVERSITY
- **Principal Investigator:** Philippe Mourrain
- **Activity code:** P50 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2023
- **Award amount:** $396,570
- **Award type:** 5
- **Project period:** 2022-09-06 → 2027-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10698080

## Citation

> US National Institutes of Health, RePORTER application 10698080, Project 4: Whole-brain and body characterization of sleep disturbances and interventions in Fmr1, Shank3 and Cntnap2 knockout zebrafish (5P50HD109861-02). Retrieved via AI Analytics 2026-05-21 from https://api.ai-analytics.org/grant/nih/10698080. Licensed CC0.

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