Overexpression screen of chromosome 21 genes reveals modulators of Sonic hedgehog signaling relevant to Down syndrome.

Trisomy 21 and mutations in the Sonic hedgehog (SHH) signaling pathway cause overlapping and pleiotropic phenotypes including cerebellar hypoplasia, craniofacial abnormalities, congenital heart defects and Hirschsprung disease. Trisomic cells derived from individuals with Down syndrome possess deficits in SHH signaling, suggesting that overexpression of human chromosome 21 genes may contribute to SHH-associated phenotypes by disrupting normal SHH signaling during development. However, chromosome 21 does not encode any known components of the canonical SHH pathway. Here, we sought to identify chromosome 21 genes that modulate SHH signaling by overexpressing 163 chromosome 21 cDNAs in a series of SHH-responsive mouse cell lines. We confirmed overexpression of trisomic candidate genes using RNA sequencing in the cerebella of Ts65Dn and TcMAC21 mice, model systems for Down syndrome. Our findings indicate that some human chromosome 21 genes, including DYRK1A, upregulate SHH signaling, whereas others, such as HMGN1, inhibit SHH signaling. Individual overexpression of four genes (B3GALT5, ETS2, HMGN1 and MIS18A) inhibits the SHH-dependent proliferation of primary granule cell precursors. Our study prioritizes dosage-sensitive chromosome 21 genes for future mechanistic studies. Identification of the genes that modulate SHH signaling may suggest new therapeutic avenues for ameliorating Down syndrome phenotypes.

Animal models of human insomnia.

Insomnia disorder (chronic sleep continuity disturbance) is a debilitating condition affecting 5%-10% of the adult population worldwide. To date, researchers have attempted to model insomnia in animals through breeding strategies that create pathologically short-sleeping individuals or with drugs and environmental contexts that directly impose sleeplessness. While these approaches have been invaluable for identifying insomnia susceptibility genes and mapping the neural networks that underpin sleep-wake regulation, they fail to capture concurrently several of the core clinical diagnostic features of insomnia disorder in humans, where sleep continuity disturbance is self-perpetuating, occurs despite adequate sleep opportunity, and is often not accompanied by significant changes in sleep duration or architecture. In the present review, we discuss these issues and then outline ways animal models can be used to develop approaches that are more ecologically valid in their recapitulation of chronic insomnia's natural aetiology and pathophysiology. Conditioning of self-generated sleep loss with these methods promises to create a better understanding of the neuroadaptations that maintain insomnia, including potentially within the infralimbic cortex, a substrate at the crossroads of threat habituation and sleep.

Sleep continuity, timing, quality, and disorder are associated with suicidal ideation and suicide attempts among college students.

Objective: To evaluate sleep continuity, timing, quality, and disorder in relation to suicidal ideation and attempts among college students. Participants: Eight hundred eighty-five undergraduates aged 18-25 in the southwestern United States. Methods: Participants completed questionnaires on sleep, suicide risk, mental health, and substance use. Differences in sleep variables were compared by lifetime and recent suicidal ideation and suicide attempts using covariate-adjusted and stepwise regression models. Results: A total of 363 (41.0%) individuals reported lifetime suicidal ideation, of whom 172 (47.4%) reported suicidal ideation in the last 3 months and 97 (26.7%) had attempted suicide in their lifetime. Sleep disturbances were prevalent among those with lifetime suicidal ideation or a lifetime suicide attempt. Insomnia was identified as the best predictor of recent suicidal ideation, but this relationship did not survive adjustment for covariates. Conclusions: Sleep continuity, quality, and sleep disorders are broadly associated with suicidal thoughts and behaviors among college students.

Social Jetlag and Other Aspects of Sleep Are Linked to Non-Suicidal Self-Injury Among College Students.

INTRODUCTION: Disrupted sleep is associated with non-suicidal self-injury (NSSI) in young adults, but many specific features of sleep continuity and timing have yet to be examined. Additionally, the psychological mechanisms linking sleep to NSSI are unclear. The present study evaluated 14 sleep variables as classifiers of lifetime or recent NSSI and examined potential confounding and mediating factors. METHODS: A sample of 885 college students provided measures of sleep continuity (e.g., duration, timing, fragmentation), nightmares, insomnia, and perceived sleep control. Lifetime and past 3-month NSSI were measured using a self-report version of the Columbia Suicide Severity Ratings Scale. Bidirectional stepwise regression identified significant sleep classifiers and subsequent models examined their associations with NSSI after adjusting for covariates and through potential psychological mediators. RESULTS: Only absolute social jetlag was associated with recent NSSI, even after adjusting for covariates, such that each additional hour difference between weekday and weekend sleep schedules was associated with a 17% greater risk of recent NSSI. Nightmares, weekend sleep efficiency, and perceived sleep control were associated with lifetime NSSI, although only weekend sleep efficiency remained associated after adjusting for covariates. Bootstrap mediations identified negative urgency as a partial mediator for recent and lifetime NSSI, and lack of premeditation and perceived burdensomeness as partial mediators for lifetime NSSI. CONCLUSIONS: The timing and consistency of young adults' sleep schedules may be of greater importance to NSSI among college students than insomnia or insufficient sleep. Future studies of sleep and NSSI should include these measures as potential risk factors. HIGHLIGHTSDifferences between weekday/weekend sleep timing are linked to recent NSSI.Negative urgency partially mediates poor sleep on recent and lifetime NSSI.Sleep shares a multifaceted relationship with NSSI risk in college students.

Sleep loss suicidal ideation: the role of trait extraversion.

Background: It is known that sleep disturbance is associated with increased suicidal thinking. Moreover, completed suicides, when adjusted for the proportion of the populace that is awake at a given time, are more probable during the late night/early morning hours. Despite these concerns, no studies have examined the role of trait-like individual differences in vulnerability to suicidal ideation during sleep deprivation or insomnia. In two separate studies, we examined whether the trait of extraversion is predictive of changes in suicidal thinking following two nights of sleep deprivation and among individuals meeting the criteria for insomnia. Methods: Study 1: Twenty-five healthy military personnel (20 males), ages 20-35 completed the NEO-PI-R Extraversion scale and the Suicidal Ideation (SUI) scale of the Personality Assessment Inventory (PAI). Participants completed 77 h of continuous sleep deprivation. After 56 h of sleep deprivation, participants completed the SUI scale a second time. We predicted a change in SUI scores from baseline extraversion. Study 2: 2,061 adults aged 18-79 (900 males) were divided into two groups based on the clinical threshold (≥ 10) on the Insomnia Severity Index (ISI) and completed measures of extraversion and depression, including the suicide item of the Patient Health Questionnaire-9 (PHQ9). Results: Study 1: After controlling for the caffeine group and changes in PAI Depression, Extraversion scores were used to predict changes in SUI scores using stepwise multiple linear regression. Higher Extraversion was significantly associated with increased non-clinical suicidal ideation following sleep loss, ß = 0.463, partial r = 0.512, p = 0.013. Study 2: After controlling for depression, the effect of insomnia on suicidal ideation was moderated by trait extraversion (p < 0.0001). Overall, the presence or absence of insomnia had little effect on individuals low in trait extraversion (i.e., introverts), but insomnia was associated with significantly higher suicidal ideation among high trait extraverted individuals. Conclusions: Higher trait extraversion was associated with increased vulnerability to suicidal ideation between rested baseline and total sleep deprivation and was associated with greater suicidal ideation among those meeting criteria for clinically severe insomnia. These findings point to a potential trait-like vulnerability factor that may further our understanding of sleep disruption in the phenomenology of suicide.

Insomnia and the Interpersonal Theory of suicide among civilians, service members, and veterans.

BACKGROUND: Insomnia is associated with suicide risk in civilian and military populations. Thwarted belongingness is proposed as a mediator of this relationship under the Interpersonal Theory of Suicide (IPTS). The present study explored how insomnia relates to suicidal ideation in conjunction with thwarted belongingness among civilians, Service members, and Veterans. METHODS: Data from the Military Suicide Research Consortium for N = 6556 individuals (6316 with non-missing suicidal ideation status) were divided into 4 subgroups: civilians, never deployed Service members, previously deployed Service members, and Veterans. Robust Poisson models evaluated the associations between insomnia severity/subtype and current suicidal ideation, with bootstrap mediation models assessing thwarted belongingness as a mediator. RESULTS: A 5-point increase in insomnia severity was associated with a 38% increased risk for current suicidal ideation among civilians, a 56% greater risk among never deployed Service members, an 83% greater risk among previously deployed Service members, and a 37% greater risk among Veterans. Moreover, active Service members showed greater associations between difficulty falling asleep and staying asleep with suicidal ideation than civilians. These associations were independent of covariates and only mediated by thwarted belongingness among Veterans. CONCLUSIONS: The relationship between insomnia and suicide is not purely explained by thwarted belongingness except among Veterans. Future research should explore additional psychological and neurobiological mechanisms connecting insomnia and suicidality.

Associations between Insomnia Symptoms and Anxiety Symptoms in Adults in a Community Sample of Southeastern Pennsylvania, USA.

Although insomnia is reliably associated with anxiety symptoms, aspects of insomnia may differentially relate to one anxiety symptom versus another. Therefore, treatment for insomnia comorbidity with anxiety might be individually tailored to optimize treatment response. Working from this hypothesis, we analyzed data from a survey of 1007 community-dwelling adults. Insomnia was measured using the Insomnia Severity Index (ISI), categorizing items as nighttime disturbances, daytime dysfunction, or self-perceived dissatisfaction. Anxiety symptoms were measured with the Generalized Anxiety Disorder 7-item questionnaire (GAD-7). Linear and binomial logistic regression were used and adjusted for covariates. Post hoc forward stepwise analyses determined which components of the insomnia contributed to individual anxiety symptoms. Significant associations between nighttime disturbance (ß = 0.88 [0.44, 1.3]), daytime dysfunction (ß = 1.30 [0.81, 1.80]), dissatisfaction (ß = 1.20 [0.60, 1.7]) and total GAD-7 score were maintained after adjusting for covariates. Nighttime disturbance was associated with excess worrying, restlessness, irritability, and fear of catastrophe. Daytime dysfunction was associated with all symptoms except for fear of catastrophe, and self-perceived dissatisfaction was associated with all symptoms except irritability. Stepwise analyses revealed that daytime dysfunction and dissatisfaction were most consistently related to anxiety symptoms.Greater attention should be paid to daytime dysfunction in patients with insomnia and anxiety, as improving daytime functioning may improve anxiety.

Reversible Suppression of Fear Memory Recall by Transient Circadian Arrhythmia.

We tested the hypothesis that a temporary period of circadian arrhythmia would transiently impair recall of an aversive memory in Siberian hamsters (Phodopus sungorus). Unlike mice or rats, circadian arrhythmia is easily induced in this species by a one-time manipulation of their ambient lighting [i.e., the disruptive phase shift (DPS) protocol]. Hamsters were conditioned to associate footshocks with a shock chamber (context) and with a predictive auditory tone (cue), and then exposed to the DPS protocol. Following DPS, animals either became arrhythmic (ARR), reentrained to the light-dark cycle (ENT), or became arrhythmic for < 14 days before their circadian locomotor rhythms spontaneously recovered and reentrained (ARR-ENT). Tests for contextual memory showed that freezing was decreased 9-10 days post-DPS when both ARR and ARR-ENT groups were arrhythmic. Once ARR-ENT animals reentrained (day 41), however, freezing was elevated back to Pre-DPS levels and did not differ from those observed in ENT hamsters. ENT animals maintained high levels of freezing at both time points, whereas, freezing remained low in ARR hamsters. In contrast to contextual responses, cued responses were unaffected by circadian arrhythmia; all three groups exhibited elevated levels of freezing in response to the tones. The differential impact of circadian arrhythmia on contextual versus cued associative memory suggests that arrhythmia preferentially impacts memory processes that depend on the hippocampus.

The Mind After Midnight: Nocturnal Wakefulness, Behavioral Dysregulation, and Psychopathology.

Sufficient sleep with minimal interruption during the circadian/biological night supports daytime cognition and emotional regulation. Conversely, disrupted sleep involving significant nocturnal wakefulness leads to cognitive and behavioral dysregulation. Most studies to-date have examined how fragmented or insufficient sleep affects next-day functioning, but recent work highlights changes in cognition and behavior that occur when someone is awake during the night. This review summarizes the evidence for day-night alterations in maladaptive behaviors, including suicide, violent crime, and substance use, and examines how mood, reward processing, and executive function differ during nocturnal wakefulness. Based on this evidence, we propose the Mind after Midnight hypothesis in which attentional biases, negative affect, altered reward processing, and prefrontal disinhibition interact to promote behavioral dysregulation and psychiatric disorders.

Sleep Deprivation Does Not Influence Photic Resetting of Circadian Activity Rhythms in Drosophila.

Previous investigations in humans and rodent animal models have assessed the interplay of sleep in the circadian system's phase responses to nighttime light exposure. The resulting data have been mixed, but generally support a modulatory role for sleep in circadian photic resetting (not an absolute requirement). Drosophila have been historically used to provide important insights in the sleep and circadian sciences. However, no experiments to date have evaluated how immediate sleep need or recent sleep history affects their pacemaker's phase readjustments to light. We did so in the current study by (1) forcing separate groups of animals to stay awake for 1 or 4 h after they were shown a broadspectrum pulse (15 min during the first half of the night, 950 lux), or (2) placing them on a restricted sleep schedule for a week before light presentation without any subsequent sleep disruption. Forced sleep restriction, whether acute or chronic, did not alter the size of light-induced phase shifts. These data are consistent with observations made in other diurnal animals and raise the possibility, more broadly, that phototherapies applied during sleep-such as may be necessary during the winter months-may still be efficacious in individuals experiencing sleep-continuity problems such as insomnia.

Investigation of the aging clock's intermittent-light responses uncovers selective deficits to green millisecond flashes.

The central pacemaker of flies, rodents, and humans generates less robust circadian output signals across normative aging. It is not well understood how changes in light sensitivity might contribute to this phenomenon. In the present study, we summarize results from an extended data series (n = 5681) showing that the locomotor activity rhythm of aged Drosophila can phase-shift normally to intermittently spaced episodes of bright polychromatic light exposure (600 lx) but that deficits emerge in response to 8, 16, and 120-millisecond flashes of narrowband blue (λm, 452 nm) and green (λm, 525 nm) LED light. For blue, phase-resetting of the activity rhythm of older flies is not as energy efficient as it is in younger flies at the fastest flash-exposures tested (8 milliseconds), suggesting there might be different floors of light duration necessary to incur photohabituation in each age group. For green, the responses of older flies are universally crippled relative to those of younger flies across the slate of protocols we tested. The difference in green flash photosensitivity is one of the most salient age-related phenotypes that has been documented in the circadian phase-shifting literature thus far. These data provide further impetus for investigations on pacemaker aging and how it might relate to changes in the circadian system's responses to particular sequences of light exposure tuned for wavelength, intensity, duration, and tempo.

Spectrophotometric properties of commercially available blue blockers across multiple lighting conditions.

Lenses that filter short-wavelength ("blue") light are commercially marketed to improve sleep and circadian health. Despite their widespread use, minimal data are available regarding their comparative efficacy in curtailing blue light exposure while maintaining visibility. Fifty commercial lenses were evaluated using five light sources: a blue LED array, a computer tablet display, an incandescent lamp, a fluorescent overhead luminaire, and sunlight. Absolute irradiance was measured at baseline and for each lens across the visual spectrum (380-780 nm), which allowed calculation of percent transmission. Transmission specificity was also calculated to determine whether light transmission was predominantly circadian-proficient (455-560 nm) or non-proficient (380-454 nm and 561-780 nm). Lenses were grouped by tint and metrics were compared between groups. Red-tinted lenses exhibited the lowest transmission of circadian-proficient light, while reflective blue lenses had the highest transmission. Orange-tinted lenses transmitted similar circadian-proficient light as red-tinted lenses but transmitted more non-circadian-proficient light, resulting in higher transmission specificity. Orange-tinted lenses had the highest transmission specificity while limiting biologically active light exposure in ordinary lighting conditions. Glasses incorporating these lenses currently have the greatest potential to support circadian sleep-wake rhythms.

Current Insights into Optimal Lighting for Promoting Sleep and Circadian Health: Brighter Days and the Importance of Sunlight in the Built Environment.

This perspective considers the possibility that daytime's intrusion into night made possible by electric lighting may not be as pernicious to sleep and circadian health as the encroachment of nighttime into day wrought by 20th century architectural practices that have left many people estranged from sunlight.

The Drosophila circadian phase response curve to light: Conservation across seasonally relevant photoperiods and anchorage to sunset.

Photic history, including the relative duration of day versus night in a 24-hour cycle, is known to influence subsequent circadian responses to light in mammals. Whether such modulation is present in Drosophila is currently unknown. To date, all photic phase-response curves (PRCs) generated from Drosophila have done so with animals housed under seasonally agnostic equatorial photoperiods with alternating 12-hour segments of light and darkness. However, the genus contains thousands of species, some of which populate high and low-latitude habitats (20-50° north or south of the Equator) where seasonal variations in the light-dark schedule are pronounced. Here, we address this disconnect by constructing the first high-resolution Drosophila seasonal atlas for light-induced circadian phase-resetting. Testing the light responses of over 4,000 Drosophila at 120 timepoints across 5 seasonally-relevant rectangular photoperiods (i.e., LD 8:16, 10:14, 12:12, 14:10, and 16:8; 24 hourly intervals surveyed in each), we determined that many aspects of the fly circadian PRC waveform are conserved with increasing daylength. Surprisingly though, irrespective of LD schedule, the start of the PRCs always remained anchored to the timing of subjective sunset, creating a differential overlap of the advance zone with the morning hours after subjective sunrise that was maximized under summer photoperiods and minimized under winter photoperiods. These data suggest that there may be differences in flies versus mammals as to how the photoperiod modulates the waveform and amplitude of the circadian PRC to light. On the other hand, they support the possibility that the lights-off transition determines the phase-positioning of photic PRCs across seasons and across species. More work is necessary to test this claim and whether it might factor into the timing of seasonal light responses in humans.

Systematic review of drugs that modify the circadian system's phase-shifting responses to light exposure.

We searched PubMed for primary research quantifying drug modification of light-induced circadian phase-shifting in rodents. This search, conducted for work published between 1960 and 2018, yielded a total of 146 papers reporting results from 901 studies. Relevant articles were those with any extractable data on phase resetting in wildtype (non-trait selected) rodents administered a drug, alongside a vehicle/control group, near or at the time of exposure. Most circadian pharmacology experiments were done using drugs thought to act directly on either the brain's central pacemaker, the suprachiasmatic nucleus (SCN), the SCN's primary relay, the retinohypothalamic tract, secondary pathways originating from the medial/dorsal raphe nuclei and intergeniculate leaflet, or the brain's sleep-arousal centers. While the neurotransmitter systems underlying these circuits were of particular interest, including those involving glutamate, gamma-aminobutyric acid, serotonin, and acetylcholine, other signaling modalities have also been assessed, including agonists and antagonists of receptors linked to dopamine, histamine, endocannabinoids, adenosine, opioids, and second-messenger pathways downstream of glutamate receptor activation. In an effort to identify drugs that unduly influence circadian responses to light, we quantified the net effects of each drug class by ratioing the size of the phase-shift observed after administration to that observed with vehicle in a given experiment. This allowed us to organize data across the literature, compare the relative efficacy of one mechanism versus another, and clarify which drugs might best suppress or potentiate phase resetting. Aggregation of the available data in this manner suggested that several candidates might be clinically relevant as auxiliary treatments to suppress ectopic light responses during shiftwork or amplify the circadian effects of timed bright light therapy. Future empirical research will be necessary to validate these possibilities.

The translational neuroscience of sleep: A contextual framework.

Sleep is entwined across many physiologic processes in the brain and periphery, thereby exerting tremendous influence on our well-being. Yet sleep exists in a social-environmental context. Contextualizing sleep health with respect to its determinants­from individual- to societal-level factors­would enable neuroscientists to more effectively translate sleep health into clinical practice. Key challenges and opportunities pertain to (i) recognizing and exploring sleep's functional roles, (ii) clarifying causal mechanisms in relation to key outcomes, (iii) developing richer model systems, (iv) linking models to known contextual factors, and (v) leveraging advances in multisensory technology. Meeting these challenges and opportunities would help transcend disciplinary boundaries such that social-environmental considerations related to sleep would become an ever-greater presence in the clinic.

Nocturnal and Morning Wakefulness Are Differentially Associated With Suicidal Ideation in a Nationally Representative Sample.

Objective: Prior studies indicate nocturnal wakefulness is associated with suicide, while morning wakefulness is linked to reduced suicidal ideation. These relationships, however, may be confounded by sociodemographic factors. Therefore, this study investigated whether timing of wakefulness was associated with suicidal ideation in a nationally representative sample.Methods: Data were collected from the US National Health and Nutrition Examination Survey for the years 2015 to 2018, resulting in a final sample of 10,166 participants (51.1% female) with complete data available on suicidal ideation status, time to bed, and time out of bed. Population-weighted logistic regression models estimated the associations between time spent out of bed (ie, being awake) and suicidal ideation.Results: A total of 385 survey participants (47.5% female) reported suicidal ideation in the past 2 weeks for a population-weighted prevalence of 3.37% (95% CI, 2.85%-3.87%). Wakefulness between 11:00 pm and 5:00 am was associated with suicidal ideation (OR = 1.16; 95% CI, 1.08-1.24 per hour), even after adjustment for sociodemographic factors and symptoms of sleep disorders, but not after adjustment for the severity of depression symptoms. Conversely, wakefulness between 5:00 am and 11:00 am was associated with reduced odds of suicidal ideation (OR = 0.77; 95% CI, 0.70-0.85 per hour) in all models.Conclusions: Individuals who spent more time awake at night were more likely to have recent suicidal ideation, while the opposite was true for those with more time spent awake in the morning. Moreover, these associations were independent of sociodemographic factors and thus not confounded by varying rates of suicidal ideation in different populations.

Forebrain Shh overexpression improves cognitive function and locomotor hyperactivity in an aneuploid mouse model of Down syndrome and its euploid littermates.

Down syndrome (DS) is the leading genetic cause of intellectual disability and causes early-onset dementia and cerebellar hypoplasia. The prevalence of attention deficit hyperactivity disorder is elevated in children with DS. The aneuploid DS mouse model "Ts65Dn" shows prominent brain phenotypes, including learning and memory deficits, cerebellar hypoplasia, and locomotor hyperactivity. Previous studies indicate that impaired Sonic hedgehog (Shh) signaling contributes to neurological phenotypes associated with DS and neurodegenerative diseases. However, because of a lack of working inducible Shh knock-in mice, brain region-specific Shh overexpression and its effects on cognitive function have not been studied in vivo. Here, with Gli1-LacZ reporter mice, we demonstrated that Ts65Dn had reduced levels of Gli1, a sensitive readout of Shh signaling, in both hippocampus and cerebellum at postnatal day 6. Through site-specific transgenesis, we generated an inducible human Shh knock-in mouse, TRE-bi-hShh-Zsgreen1 (TRE-hShh), simultaneously expressing dually-lipidated Shh-Np and Zsgreen1 marker in the presence of transactivator (tTA). Double transgenic mice "Camk2a-tTA;TRE-hShh" and "Pcp2-tTA;TRE-hShh" induced Shh overexpression and activated Shh signaling in a forebrain and cerebellum, respectively, specific manner from the perinatal period. Camk2a-tTA;TRE-hShh normalized locomotor hyperactivity and improved learning and memory in 3-month-old Ts65Dn, mitigated early-onset severe cognitive impairment in 7-month-old Ts65Dn, and enhanced spatial cognition in euploid mice. Camk2a-tTA;TRE-hShh cohort maintained until 600days old showed that chronic overexpression of Shh in forebrain from the perinatal period had no effect on longevity of euploid or Ts65Dn. Pcp2-tTA;TRE-hShh did not affect cognition but mitigated the phenotype of cerebellar hypoplasia in Ts65Dn. Our study provides the first in vivo evidence that Shh overexpression from the perinatal period protects DS brain integrity and enhances learning and memory in normal mice, indicating the broad therapeutic potential of Shh ligand for other neurological conditions. Moreover, the first inducible hShh site-specific knock-in mouse could be widely used for spatiotemporal Shh signaling regulation.