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Bastien, B. L.

Publications and source records attributed to Bastien, B. L..

5 recordsLinked to original sources

Znf804a is a regulator of circadian behaviors in zebrafish

Sleep disturbances are common among individuals with schizophrenia and can exacerbate disruptions in cognitive processes like learning and memory. Elucidating pharmacologically targetable molecular pathways perturbed by schizophrenia genes may uncover new treatment avenues. Here, we investigated the relationship of the schizophrenia-associated gene znf804a with sleep and circadian pathways. Using multi-day behavior tracking, we showed that znf804a zebrafish mutants displayed changes in sleep and circadian behaviors when light cues were removed. Through bulk RNA sequencing of fish raised under normal light cycling and dark-only conditions, we identified altered gene expression in the core and auxiliary pathways controlling circadian rhythms. Expression of fbxl3a, which encodes a modulator of the core negative feedback regulator of the clock, decreased in a dose-dependent manner as znf804a mutant copy number increased. Further analysis also revealed shifts in the relative abundance of specific transcripts, including idh1, suggesting znf804a could influence transcript processing or stability. Together, these findings link a ZNF804A ortholog to sleep and circadian behaviors and identify the regulation of fbxl3a and transcript processing as candidate mechanisms through which this schizophrenia risk gene may influence circadian biology.

genetics↗

Zebrafish screen of schizophrenia risk genes reveals convergent dysregulation of cholesterol metabolism

Rare coding variants provide a tractable entry point for understanding the molecular mechanisms underlying schizophrenia risk. Here, we generated and characterized zebrafish lines with mutations in the orthologs of >20 human schizophrenia-associated genes, including eight of the top ten SCHEMA genes, genes disrupted in childhood-onset schizophrenia (COS), and genes located within recurrent copy number variants. Whole-brain phospho-Erk activity mapping and behavioral profiling identified phenotypes in multiple mutant lines. We prioritized a protein-truncating mutation in sp4, which encodes an activity-dependent transcription factor, and a COS-associated missense mutation in atp1a3a, which encodes a Na+/K+ ATPase pump, for additional characterization. Both knockout and point mutations in atp1a3a disrupted brain activity and behavior in larvae and impaired navigation of a Y-maze in juveniles. Bulk RNA sequencing data from adult sp4 and atp1a3a brains highlighted convergent upregulation of sterol biosynthesis pathways, including increased expression of srebf2 and msmo1. Analysis of previously published telencephalon single-cell data demonstrated that cholesterol synthesis genes are enriched in astrocyte-like cells and increase in expression during post-larval development. Consistent with transcriptomic findings, filipin staining indicated increased free cholesterol in juvenile sp4 and atp1a3a mutant brains. Our findings identify dysregulation of glial and sterol-associated programs as a shared molecular consequence of two distinct schizophrenia risk mutations. Although whether sterol pathway dysregulation represents a primary pathogenic mechanism or a secondary response to changes in neuronal activity requires further investigation, the convergence observed between genetic models and developmental stages suggests that disruptions to lipid homeostasis could represent a shared feature of schizophrenia disease biology.

genetics↗

Allele-specific rescue of neurexin behavioral phenotypes by monoamine-targeting compounds

Neurexins are synaptic adhesion molecules associated with neurologic changes in humans, including neurodevelopmental delay, autism, schizophrenia, Tourette syndrome, and seizures. The NRXN1 gene produces >100 protein isoforms through alternative promoters and extensive splicing, which are differentially impacted by NRXN1 variants found in patients. Yet pharmacologic targeting of NRXN1 isoforms or deletions has not been comprehensively studied. Here, we developed a behavioral screening approach in C. elegans to identify small molecule compounds that modify the decreased activity levels caused by isoform-specific deletions of neurexin(nrx-1). Screening 190 compounds, we discovered that monoamine-targeting compounds differentially improve behavioral phenotypes depending on which nrx-1 isoforms are disrupted. Broad modulation of monoamine signaling, or antagonism of specific serotonin receptors, are required to increase the activity of both alleles tested. The FDA-approved atypical antipsychotic olanzapine was the sole validated compound achieving Z-scores >2 in both screens, which notably also rescued behavioral phenotypes of C. elegans harboring a conserved autism-associated NRXN1 missense variant (L18Q/L16Q) identified in human patients. In Drosophila Nrx-1 mutants, olanzapine uniquely and significantly improved activity deficits and extended survival, demonstrating evolutionary conservation of our findings. Multi-behavior testing revealed pharmacological specificity: olanzapine improved both activity and social feeding phenotypes of nrx-1 alleles, while asenapine maleate improved activity, but worsened social feeding, indicating distinct impacts across behavioral domains. Our findings establish monoamine modulation as a conserved compensatory mechanism for neurexin loss, identify olanzapine as a lead compound for targeting neurexin loss, and demonstrate that allele stratification and pharmacogenomic approaches are needed for precision intervention in behavioral conditions. Neurexins are synaptic adhesion molecules implicated in autism, schizophrenia, and neurodevelopmental disorders. NRXN1 produces over 100 isoforms differentially affected by patient variants, yet pharmacologic targeting has not been systematically studied. We developed a C. elegans behavioral screen to identify compounds rescuing activity deficits caused by isoform-specific deletions. Screening 190 compounds, we discovered that monoamine-targeting drugs differentially improved behavioral phenotypes depending on which isoforms were disrupted. The FDA-approved antipsychotic olanzapine uniquely achieved robust rescue (Z-scores >2) across all genetic backgrounds tested, including a conserved autism-associated NRXN1 missense variant (L18Q/L16Q) identified in patients. Olanzapines efficacy was conserved in Drosophila Nrx-1 mutants, improving activity and extending survival. Multi-behavior testing revealed pharmacological specificity: olanzapine rescued both activity and social feeding phenotypes, while asenapine differentially affected behavioral domains. Our findings establish monoamine modulation as a compensatory mechanism for neurexin loss and identify olanzapine as a therapeutic lead for precision intervention in neurexin-associated disorders.

neuroscience↗

StrIPETrack: a real-time, ROI-flexible tracking platform for high-throughput zebrafish behavior

Quantitative phenotyping is essential to studies of animal behavior, enabling systematic analysis of variation arising from natural diversity or experimental manipulation. High-throughput behavioral assays that can simultaneously test multiple animals support sufficiently powered studies of behavioral variation, but accurate tracking of each animal is critical. Furthermore, behavioral tasks and experimental arenas span a wide range of complexity, from the reaction of a single larval zebrafish to an acoustic stimulus to associative conditioning in cue-rich environments. Here, we developed and validated StrIPETrack (Structural similarity-based Image Processing for Estimation and Tracking), a Python-based, modular animal tracking software designed for flexible region-of-interest (ROI) definitions and extensibility across assays. We show that StrIPETrack measures activity comparably to our previous LabVIEW-based zebrafish tracking software and detects similar behavioral differences between wild-type clutches. In addition, StrIPETrack accurately captures behavior in a complex arena: the Y-maze. Our approach for analyzing Y-maze navigation yields an expanded set of metrics beyond turn count and direction, revealing more subtle behavioral variation. Overall, this versatile software can be applied to monitor the activity of multiple animals in parallel in both simple high-throughput and more complex assays, and can be readily adapted to new paradigms. SummaryOur open-source tracking software provides rich behavioral phenotyping of animals in many behavioral tasks. The flexible ROI design and live tracking makes the software adaptable to diverse paradigms.

animal behavior and cognition↗

Insertion of rare autism variants in synaptic genes induce novel behavioral phenotypes in C. elegans

Neurodevelopmental conditions and disorders, including autism, involve a complex interplay of genetic, environmental, and developmental factors. Despite this complexity, genetic studies have identified more than 150 candidate genes that increase risk for autism and related neurodevelopmental and neuropsychiatric conditions. Unsurprisingly, synaptic genes are a large proportion of these genes, likely due to their roles in the formation and maintenance of synaptic architecture, function, and the plasticity of neurons and circuits. The association of synaptic genes with autism and similar conditions is driven by all types of genetic variation, including inherited and de novo rare variants that have unknown impacts on the function of the gene. Here we insert 4 conserved rare variants in the C. elegans orthologs of NLGN4X, NRXN1, and SHANK3, and define their impact on gene function compared to known loss of function variants using behavioral assays. We find that the rare variants impact multiple foraging behaviors, with each gene and variant having a unique pattern of behavioral changes and functional impact. The NLGN4X(A283T) variant induced clear loss of function, while NLGN4X(G84R) induces a loss of function in one behavior, but a gain of function in another behavior. The NRXN1(L18Q) variant induced remarkable loss and gain of functions with distinct impacts across each behavior. The SHANK3(L143P) variant induced partial loss of function in a single behavior. We also identify for the first time that loss of function of shn-1/SHANK3 alters social feeding and food response behaviors. We uncover a remarkably complex impact of rare variants in synaptic genes, with differential impacts across behaviors, highlighting the importance of broad behavioral analysis and the nuanced effects of missense variants compared to loss of function alleles. Together, we define the complex functional impact of each variant on gene function, compare the impact of variants and genes across multiple behaviors, and provide further support for the use of C. elegans to define the impact of genetic variation derived from human neurodevelopmental and neuropsychiatric disorders.

genetics↗