Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2022 Aug 29;13(1):35.
doi: 10.1186/s13229-022-00514-5.

Early life sleep disruption potentiates lasting sex-specific changes in behavior in genetically vulnerable Shank3 heterozygous autism model mice

Julia S Lord  1 , Sean M Gay  1 , Kathryn M Harper  2   3 , Viktoriya D Nikolova  2   3 , Kirsten M Smith  1 , Sheryl S Moy  4   5 , Graham H Diering  6   7
Affiliations

Early life sleep disruption potentiates lasting sex-specific changes in behavior in genetically vulnerable Shank3 heterozygous autism model mice

Julia S Lord et al. Mol Autism. .

Abstract

Background: Patients with autism spectrum disorder (ASD) experience high rates of sleep disruption beginning early in life; however, the developmental consequences of this disruption are not understood. We examined sleep behavior and the consequences of sleep disruption in developing mice bearing C-terminal truncation mutation in the high-confidence ASD risk gene SHANK3 (Shank3ΔC). We hypothesized that sleep disruption may be an early sign of developmental divergence, and that clinically relevant Shank3WT/ΔC mice may be at increased risk of lasting deleterious outcomes following early life sleep disruption.

Methods: We recorded sleep behavior in developing Shank3ΔC/ΔC, Shank3WT/ΔC, and wild-type siblings of both sexes using a noninvasive home-cage monitoring system. Separately, litters of Shank3WT/ΔC and wild-type littermates were exposed to automated mechanical sleep disruption for 7 days prior to weaning (early life sleep disruption: ELSD) or post-adolescence (PASD) or undisturbed control (CON) conditions. All groups underwent standard behavioral testing as adults.

Results: Male and female Shank3ΔC/ΔC mice slept significantly less than wild-type and Shank3WT/ΔC siblings shortly after weaning, with increasing sleep fragmentation in adolescence, indicating that sleep disruption has a developmental onset in this ASD model. ELSD treatment interacted with genetic vulnerability in Shank3WT/ΔC mice, resulting in lasting, sex-specific changes in behavior, whereas wild-type siblings were largely resilient to these effects. Male ELSD Shank3WT/ΔC subjects demonstrated significant changes in sociability, sensory processing, and locomotion, while female ELSD Shank3WT/ΔC subjects had a significant reduction in risk aversion. CON Shank3WT/ΔC mice, PASD mice, and all wild-type mice demonstrated typical behavioral responses in most tests.

Limitations: This study tested the interaction between developmental sleep disruption and genetic vulnerability using a single ASD mouse model: Shank3ΔC (deletion of exon 21). The broader implications of this work should be supported by additional studies using ASD model mice with distinct genetic vulnerabilities.

Conclusion: Our study shows that sleep disruption during sensitive periods of early life interacts with underlying genetic vulnerability to drive lasting and sex-specific changes in behavior. As individuals progress through maturation, they gain resilience to the lasting effects of sleep disruption. This work highlights developmental sleep disruption as an important vulnerability in ASD susceptibility.

Keywords: Autism spectrum disorder; Brain development; Shank3; Sleep; Social behavior.

PubMed Disclaimer

Conflict of interest statement

The authors declare no competing interests.

Figures

Fig. 1
Fig. 1
Developmental sleep disruption in Shank3ΔC ASD mouse models. A-D Noninvasive home-cage sleep recordings were conducted for male and female Shank3WT/ΔC heterozygotes, Shank3ΔC/ΔC homozygotes, and wild-type (WT) littermates, at two developmental time points: juvenile (P23–P41) and adolescent (P42–P56). A Juvenile males, B juvenile females, C adolescent males, D adolescent females. A–D Left panel: trace of daily average hourly percent sleep, dark phase indicated by gray box. Center panel: average hourly percent sleep for the light or dark phase [12 h], or the daily average. Right panel: average sleep bout length observed in the light or dark phase. Genotype differences found with post hoc Tukey correction are indicated. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 (1-way ANOVA with Tukey’s test for multiple comparisons). Error bars indicate ± SEM in sleep traces; box plots show median, range, and 1st/3rd quartiles for sleep values. Statistics analysis is summarized in Additional file 1
Fig. 2
Fig. 2
Early life or post-adolescent sleep disruption. A Experimental design. Cohorts of mice undergo 7 days of sleep disruption either from P14–P21: early life sleep disruption (ELSD), or from P56–P63: post-adolescent sleep disruption (PASD). Control cohorts are left undisturbed. Control, ELSD, and PASD cohorts include male and female WT and Shank3WT/ΔC heterozygous littermates. Upon reaching adulthood (P70), control, ELSD, and PASD cohorts undergo a panel of behavioral testing lasting 6–7 weeks. B Control and ELSD pups were weighed daily from P14–P21. No differences were seen between groups (2-way ANOVA). N = 37 control pups; 39 ELSD pups. C Separate cohorts of control and ELSD treatment were killed at P21, and serum corticosterone (cort.) was measured using ELISA. No differences were seen between groups (unpaired t test). N  = 17 control, 33 ELSD. Error bars indicate ± SEM. See also Additional file 3
Fig. 3
Fig. 3
ELSD drives sex-specific changes in certain non-social behaviors in Shank3WT/ΔC heterozygotes. A–D Percent open arm time and entries for WT and Shank3WT/ΔC heterozygotes (HET) from control, ELSD, and PASD treatment groups. A Male % time spent in the open arms. B Male % open arm entries. No differences were detected in any male groups. C Female % time spent in the open arms. ELSD HET females spend significantly more time in the open arms than WT littermates. D Female % open arms entries. **P < 0.01 (unpaired t test for genotype with Tukey’s multiple comparisons test). E–F Total distance traveled in the open-field task for WT and HET males E or females F. ELSD HET males show hypo-activity in the open field test. G–H Percent pre-pulse inhibition (PPI) of acoustic startle responses for WT and HET males G or females H. ELSD HET males show reductions in PPI response. **P < 0.01 (Open field, unpaired t test with Tukey’s multiple comparisons; PPI, 2-way ANOVA with Šídák’s multiple comparisons test). Error bars indicate ± SEM. N = 8–12 mice per group/sex/genotype. Summary of statistical analysis is shown in Additional file 2. See also Additional file 4 (anxiety-like measures in open-field test)
Fig. 4
Fig. 4
ELSD/PASD treatment did not result in any lasting changes in the accelerating rotarod, marble burying, or hidden food assays. A, B Latency to fall in the accelerating rotarod assay of motor coordination and motor learning in control, ELSD, and PASD treated WT and Shank3WT/ΔC heterozygous (HET) males (A) and females (B). No differences were observed (2-way ANOVA with Šídák’s multiple comparisons test). C, D Marbles buried by control, ELSD, and PASD treated males (C) and females (D). No differences were observed (unpaired t tests with Holm–Šídák correction). E, F Latency to find hidden food in the olfaction test. No differences were observed (unpaired t tests with Holm–Šídák correction). N = 8–12 per treatment/sex/genotype
Fig. 5
Fig. 5
Social approach and social novelty preference behaviors are differentially sensitive to sex, Shank3 genotype, and sleep disruption treatments. A Examples of individual WT and Shank3WT/ΔC heterozygotes (HET) from control, ELSD, and PASD treatment groups performing the 3-chamber social approach task. Heat map indicates time in location. Mice can freely move between 3-chambers and choose to spend time engaging with the non-social stimulus (N) or with a social stimulus (S). B, C Social approach behavior: time spent in proximity with the non-social (N) and social stimulus (S) for WT and HET males (B) or females (C). ELSD HET males show no preference for the social stimulus. D, E Social novelty preference: time spent in proximity with familiar stranger 1 (S1) and novel stranger 2 (S2) for WT and HET males (D) or females (E). PASD HET males and WT females show no preference for S2. HET females show now preference for S2 under any condition tested. *P < 0.05, **P < 0.01, **P < 0.001 (paired t test with Holm–Šídák correction). N = 8–12 per treatment/sex/genotype. Summary of statistical analysis is shown in Additional file 2
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Mazzone L, Postorino V, Siracusano M, Riccioni A, Curatolo P. The relationship between sleep problems, neurobiological alterations, core symptoms of autism spectrum disorder, and psychiatric comorbidities. J Clin Med. 2018;7(5):102. doi: 10.3390/jcm7050102. - DOI - PMC - PubMed
    1. Missig G, McDougle CJ, Carlezon WA., Jr Sleep as a translationally-relevant endpoint in studies of autism spectrum disorder (ASD) Neuropsychopharmacol. 2019;45(1):90–103. doi: 10.1038/s41386-019-0409-5. - DOI - PMC - PubMed
    1. Veatch OJ, Sutcliffe JS, Warren ZE, Keenan BT, Potter MH, Malow BA. Shorter sleep duration is associated with social impairment and comorbidities in ASD. Autism Res. 2017;10(7):1221–38. doi: 10.1002/aur.1765. - DOI - PMC - PubMed
    1. MacDuffie KE, Munson J, Greenson J, Ward TM, Rogers SJ, Dawson G, et al. Sleep problems and trajectories of restricted and repetitive behaviors in children with neurodevelopmental disabilities. J Autism Dev Disord. 2020;50(11):3844–56. doi: 10.1007/s10803-020-04438-y. - DOI - PMC - PubMed
    1. MacDuffie KE, Shen MD, Dager SR, Styner MA, Kim SH, Paterson S, et al. Sleep onset problems and subcortical development in infants later diagnosed with autism spectrum disorder. Am J Psychiatry. 2020;177(6):518–25. doi: 10.1176/appi.ajp.2019.19060666. - DOI - PMC - PubMed

Publication types