A fluid-walled microfluidic platform for human neuron microcircuits and directed axotomy.

In our brains, different neurons make appropriate connections; however, there remain few in vitro models of such circuits. We use an open microfluidic approach to build and study neuronal circuits in vitro in ways that fit easily into existing bio-medical workflows. Dumbbell-shaped circuits are built in minutes in standard Petri dishes; the aqueous phase is confined by fluid walls - interfaces between cell-growth medium and an immiscible fluorocarbon, FC40. Conditions are established that ensure post-mitotic neurons derived from human induced pluripotent stem cells (iPSCs) plated in one chamber of a dumbbell remain where deposited. After seeding cortical neurons on one side, axons grow through the connecting conduit to ramify amongst striatal neurons on the other - an arrangement mimicking unidirectional cortico-striatal connectivity. We also develop a moderate-throughput non-contact axotomy assay. Cortical axons in conduits are severed by a media jet; then, brain-derived neurotrophic factor and striatal neurons in distal chambers promote axon regeneration. As additional conduits and chambers are easily added, this opens up the possibility of mimicking complex neuronal networks, and screening drugs for their effects on connectivity.

Early deficits in an in vitro striatal microcircuit model carrying the Parkinson's GBA-N370S mutation.

Understanding medium spiny neuron (MSN) physiology is essential to understand motor impairments in Parkinson's disease (PD) given the architecture of the basal ganglia. Here, we developed a custom three-chambered microfluidic platform and established a cortico-striato-nigral microcircuit partially recapitulating the striatal presynaptic landscape in vitro using induced pluripotent stem cell (iPSC)-derived neurons. We found that, cortical glutamatergic projections facilitated MSN synaptic activity, and dopaminergic transmission enhanced maturation of MSNs in vitro. Replacement of wild-type iPSC-derived dopamine neurons (iPSC-DaNs) in the striatal microcircuit with those carrying the PD-related GBA-N370S mutation led to a depolarisation of resting membrane potential and an increase in rheobase in iPSC-MSNs, as well as a reduction in both voltage-gated sodium and potassium currents. Such deficits were resolved in late microcircuit cultures, and could be reversed in younger cultures with antagonism of protein kinase A activity in iPSC-MSNs. Taken together, our results highlight the unique utility of modelling striatal neurons in a modular physiological circuit to reveal mechanistic insights into GBA1 mutations in PD.

The link between adolescent girls' interpersonal emotion regulation with parents and peers and depressive symptoms: A real-time investigation.

Adolescents often experience heightened socioemotional sensitivity warranting their use of regulatory strategies. Yet, little is known about how key socializing agents help regulate teens' negative emotions in daily life and implications for long-term adjustment. We examined adolescent girls' interpersonal emotion regulation (IER) with parents and peers in response to negative social interactions, defined as parent and peer involvement in the teen's enactment of emotion regulation strategies. We also tested associations between rates of daily parental and peer IER and depressive symptoms, concurrently and one year later. Adolescent girls (N = 112; Mage = 12.39) at temperamental risk for depressive disorders completed a 16-day ecological momentary assessment protocol measuring reactivity to negative social interactions, parental and peer IER, and current negative affect. Results indicated that adolescents used more adaptive strategies with peers and more maladaptive strategies with parents in daily life. Both parental and peer IER down-regulated negative affect, reflected by girls' decreased likelihood of experiencing continued negative affect. Higher proportions of parental adaptive IER predicted reduced depressive symptoms one year later. Findings suggest that both parents and peers effectively help adolescent girls down-regulate everyday negative emotions; however, parents may offer more enduring benefits for long-term adjustment.

Daily manifestations of caregiver- and self-reported maladaptive personality traits in adolescent girls.

Establishing maladaptive personality traits at a younger age in a developmentally appropriate and clinically tangible way may alert clinicians to dysfunction earlier, and thus reduce the risk of significant impairment later in life. The fifth edition of the Diagnostic and Statistical Manual of Mental Disorders (DSM-5) Alternative Model for Personality Disorders (AMPD) provides a set of traits useful for organizing behavioral and experiential patterns central to daily personality functioning. The goal of the present study was to evaluate manifestations indicative of AMPD traits via ambulatory assessments in the daily lives of adolescent girls. Caregivers and girls (N = 129; age: M = 12.27, SD = 0.80) provided baseline assessments of girls' trait vulnerabilities (negative affectivity, detachment, antagonism, disinhibition, psychoticism) and girls additionally completed a 16-day ecological momentary assessment protocol (N = 5,036 observations), rating social behaviors and experiences in their daily lives. Multilevel structural equation models revealed that trait vulnerabilities were linked to more extreme shifts in interpersonal experiences and behaviors from one moment to the next, suggesting that maladaptive personality traits were linked to greater variability. Furthermore, AMPD traits were positively and strongly related to negative affect in daily interpersonal situations. More specifically, girls' trait ratings were associated with elevated mean-levels of boredom, as well as interpersonal tension. Caregiver-reports complemented this perspective of dissatisfying social interactions, suggesting that especially detachment and antagonism accounted for lower levels of social connectedness and more variability in social activities in girls' daily lives. Results are discussed in terms of the short-term dynamics and related intervention targets of developmental personality pathology. (PsycInfo Database Record (c) 2023 APA, all rights reserved).

The role of anxiety and gender in anticipation and avoidance of naturalistic anxiety-provoking experiences during adolescence: An ecological momentary assessment study.

Objective: Anxiety symptoms often increase in late childhood/early adolescence, particularly among girls. However, few studies examine anxiety-relevant gender differences during anticipation and avoidance of naturalistic experiences during adolescence. The current study uses ecological momentary assessment (EMA) to examine associations among clinical anxiety, gender, anticipation, and attempted avoidance of person-specific anxiety-provoking experiences in youth ages 8-18. Method: 124 youth (73 girls) completed 7 consecutive days of EMA. Seventy participants (42 girls) met criteria for one or more anxiety disorders, while the remaining 54 were healthy controls (31 girls). Participants reported the experience that they were "most worried about happening that day" and completed ratings about that event including whether they attempted to avoid that experience. Multilevel models examined whether diagnostic group (anxious, healthy), gender (boys, girls), or their interaction predicted anticipatory ratings or avoidance of these experiences. Results: Analyses revealed significant diagnostic group by gender interactions for anticipatory ratings. Specifically, anxious girls reported greater worry and predicted more negative outcomes related to future experiences. However, only a main effect of diagnostic group emerged for attempted avoidance. Finally, anticipatory worry predicted higher rates of attempted avoidance, but this association did not vary by diagnostic group, gender, or their interaction. Conclusion: These findings extend the literature on the interplay of anticipation and avoidance to person-specific naturalistic experiences in pediatric anxiety. They reveal that anxious girls report more anticipatory anxiety and worry, while avoidance of real-world anxiety-provoking scenarios is a key concern for anxious youth independent of gender. By using EMA to examine person-specific anxiety-inducing experiences we can begin to understand how these processes and experiences unfold in the real world.

Storm Clouds and Silver Linings: Day-to-Day Life in COVID-19 Lockdown and Emotional Health in Adolescent Girls.

OBJECTIVE: We examined risk and protective factors for emotional health problems in adolescent girls during the COVID-19 pandemic. We investigated pre- to early-pandemic changes in symptoms of anxiety and depression, documented daily activities and perceived positive and negative impacts of the pandemic, and linked perceived positive and negative impacts of the pandemic to real-time changes in emotional health. METHODS: The study was a 10-day daily diary study with 93 U.S. adolescent girls (aged 12-17; 68% White non-Hispanic) at temperamental risk for anxiety and depression, conducted in April/May 2020 when all participants were under state-issued stay-at-home orders. Girls provided daily reports of positive and negative affect, depressive and anxious symptoms, activities, and positive and negative impacts resulting from the pandemic. RESULTS: Girls reported engaging in many activities that may contribute to well-being. Mixed effects analyses revealed positive impacts associated with improved same-day emotional health such as more time for family and relaxation and reduced pressure from school/activities. Negative impacts associated with poorer same-day emotional health included problems with online schooling, lack of space/privacy, lack of a regular schedule, and family conflict. CONCLUSION: Findings highlight the importance of providing in-person or quality online schooling, resources and space for learning, promoting daily routines, and spending time with teens while reducing family conflict. The pandemic also appears to have offered many girls a respite from the chronic stress of modern teen life, with time to relax and engage in creative and healthy pursuits showing benefits for daily emotional health, which should be considered following the return to normal life.

Where it Hurts the Most: Peer Interactions on Social Media and in Person are Differentially Associated with Emotional Reactivity and Sustained Affect Among Adolescent Girls.

Social media (SM) use has increasingly changed how adolescents interact with their peers, yet it remains unclear how peer interactions on social media differ from in-person peer interactions. The current study evaluated whether the context (social media or in-person) of adolescent girls' worst and best peer interactions influenced their emotional responses to peer interactions and sustained affect in everyday life. In this study, a total of 110 adolescent girls (11-13 years old; mean age = 12.28 years) completed ecological momentary assessment (EMA) for 16 days following an initial baseline visit. Participants reported their worst (i.e., most negative) and best (i.e., most positive) interactions with peers since the last prompt, the context in which it occurred (social media or in-person), emotional reactivity during the interaction, and momentary affect. Multilevel models indicated that negative peer interactions that occurred on social media were more likely to be associated with sustained negative affect, but not negative emotional reactivity during the interaction. Positive interactions on social media were more likely to be associated with both lower positive emotional reactivity and lower sustained positive affect. Findings indicate that peer interactions on social media may differentially impact girls' emotional reactivity and sustained affect, particularly for positive interactions with peers. Findings highlight that social media and in-person peer interactions may impact how girls experience and respond to positive and negative peer interactions, which may have implications for peer relationships and onset of psychopathology during this vulnerable period.

Sleep and suicide: A systematic review and meta-analysis of longitudinal studies.

The current review provides a quantitative synthesis of the empirical literature on sleep disturbance as a risk factor for suicidal thoughts and behaviors (STBs). A systematic search of PsycINFO, MEDLINE, and the references of prior reviews resulted in 41 eligible studies included in this meta-analysis. Sleep disturbance, including insomnia, prospectively predicted STBs, yielding small-to-medium to medium effect sizes for these associations. Complicating interpretation of these findings however, is that few studies of suicidal ideation and suicide attempts, as well as none of suicide deaths, assessed short-term risk (i.e., employed follow-up assessments of under a month). Such studies are needed to evaluate current conceptualizations of sleep dysregulation as being involved in acute risk for suicidal behavior. This want of short-term risk studies also suggests that current clinical recommendations to monitor sleep as a potential warning sign of suicide risk has a relatively modest empirical basis, being largely driven by cross-sectional or retrospective research. The current review ends with recommendations for generating future research on short-term risk and greater differentiation between acute and chronic aspects of sleep disturbance, and by providing a model of how sleep disturbance may confer risk for STBs through neuroinflammatory and stress processes and associated impairments in executive control.

Advancing clinical neuroscience through enhanced tools: Pediatric social anxiety as an example.

BACKGROUND: Clinical researchers face challenges when trying to quantify diverse processes engaged during social interactions. We report results from two studies, each demonstrating the potential utility of tools for examining processes engaged during social interactions. METHOD: In the first study, youth (n = 57) used a smartphone-based tool to rate mood and responses to social events. A subset (n = 20) completed the second, functional magnetic resonance imaging study. This second study related anxiety to error-evoked brain responses in two social conditions-while being observed and when alone. We also combined these tools to bridge clinical, social-contextual, and neural levels of measurement. RESULTS: Results from the first study showed an association between negatively-perceived social experiences and a range of negative emotions. In the second study there was a positive correlation during error monitoring between social-anxiety severity and context-specific activation of the pregenual anterior cingulate cortex. Finally, during imaging, the perceived quality of peer interactions as assessed using the smartphone-based tool, interacted with social context to predict levels of activation in the hippocampus and superior frontal gyrus. CONCLUSIONS: By improving measurement, enhanced tools may provide new means for studying relationships among anxiety, brain function, and social interactions.

Transgenic Strategies for Sparse but Strong Expression of Genetically Encoded Voltage and Calcium Indicators.

Rapidly progressing development of optogenetic tools, particularly genetically encoded optical indicators, enables monitoring activities of neuronal circuits of identified cell populations in longitudinal in vivo studies. Recently developed advanced transgenic approaches achieve high levels of indicator expression. However, targeting non-sparse cell populations leads to dense expression patterns such that optical signals from neuronal processes cannot be allocated to individual neurons. This issue is particularly pertinent for the use of genetically encoded voltage indicators whose membrane-delimited signals arise largely from the neuropil where dendritic and axonal membranes of many cells intermingle. Here we address this need for sparse but strong expression of genetically encoded optical indicators using a titratable recombination-activated transgene transcription to achieve a Golgi staining-type indicator expression pattern in vivo. Using different transgenic strategies, we also illustrate that co-expression of genetically encoded voltage and calcium indicators can be achieved in vivo for studying neuronal circuit input-output relationships.