Removal of developmentally regulated microexons has a minimal impact on larval zebrafish brain morphology and function DOI Open Access

Caleb CS Calhoun,

Mary E.S. Capps,

Kristie Muya

и другие.

Опубликована: Дек. 24, 2024

Microexon splicing is a vertebrate-conserved process through which small, often in-frame, exons are differentially included during brain development and across neuron types. Although the protein sequences encoded by these highly conserved can mediate interactions, neurobiological functions of only small number have been characterized. To establish more generalized understanding their roles in development, we used CRISPR/Cas9 to remove 45 microexons zebrafish assessed larval activity, morphology, behavior. Most mutants had minimal or no phenotypes at this developmental stage. Among previously studied microexons, uncovered baseline stimulus-driven for two (meA meB) ptprd reduced activity telencephalon tenm3 B 0 isoform. mild, neural were discovered several that not characterized, including ppp6r3, sptan1, dop1a, rapgef2, dctn4, vti1a , meaf6 . This study establishes general approach investigating alternative events prioritizes downstream analysis.

Язык: Английский

Genetic context of transgene insertion can influence neurodevelopment in zebrafish DOI Creative Commons
Anna J. Moyer,

Alexia Barcus,

Mary E.S. Capps

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2025, Номер unknown

Опубликована: Март 5, 2025

ABSTRACT The Gal4/ UAS system is used across model organisms to overexpress target genes in precise cell types and relies on generating transgenic Gal4 driver lines. In zebrafish, the Tg(elavl3:KalTA4) ( HuC ) line drives robust expression neurons. We observed an increased prevalence of swim bladder defects zebrafish larvae compared wildtype siblings, which prompted us investigate whether display additional neurobehavioral phenotypes. showed alterations brain activity, morphology, behavior, including hindbrain size reduced activity cerebellum. Bulk RNA-seq analysis revealed dysregulation transcriptome suggested ratio neuronal progenitor cells differentiated To understand these phenotypes derive from toxicity or positional effects related transgenesis, we economical low-pass whole genome sequencing map Tol2- mediated insertion site chromosome eight. Reduced neighboring gene gadd45ga , a known cycle regulator, consistent with proliferation suggests role for effects. Challenges creating alternative pan-neuronal lines include length elavl3 promoter (over 8 kb) random using traditional transgenesis methods. facilitate generation lines, cloned five promoters atp6v0cb smaller elavl3, rtn1a, sncb stmn1b ranging 1.7 kb 4.3 created KalTA4 Tol2 phiC31 integrase-based pIGLET system. Our study highlights importance appropriate genetic controls interrogating potential new ARTICLE SUMMARY tool that allows researchers specific by pairing “driver” “reporter” line. more than 500 have been generated methods insert DNA into sites genome. Here, authors report line, has phenotypes, changes structure, behavior. These findings emphasize highlight utility targeted transgenesis.

Язык: Английский

Процитировано

0
Dynamic convergence of autism disorder risk genes across neurodevelopment DOI Creative Commons
Meilín Fernández García, Kayla G. Townsley, April Pruitt

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Авг. 24, 2024

Over a hundred risk genes underlie for autism spectrum disorder (ASD) but the extent to which they converge on shared downstream targets increase ASD is unknown. To test hypothesis that cellular context impacts nature of convergence, here we apply pooled CRISPR approach target 29 loss-of-function in human induced pluripotent stem cell (hiPSC)-derived neural progenitor cells, glutamatergic neurons, and GABAergic neurons. Two distinct approaches (gene-level network-level analyses) demonstrate convergence greatest mature Convergent effects are dynamic, varying strength, composition, biological role between types, increasing with functional similarity examined, driven by cell-type-specific gene co-expression patterns. Stratification yield targeted drug predictions capable reversing gene-specific convergent signatures cells ASD-related behaviors zebrafish. Altogether, networks represent novel points individualized therapeutic intervention.

Язык: Английский

Процитировано

2
Removal of developmentally regulated microexons has a minimal impact on larval zebrafish brain morphology and function DOI Creative Commons

Caleb CS Calhoun,

Mary E.S. Capps,

Kristie Muya

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Авг. 20, 2024

Abstract Microexon splicing is a vertebrate-conserved process through which small, often in-frame, exons are differentially included during brain development and across neuron types. Although the protein sequences encoded by these highly conserved can mediate interactions, neurobiological functions of only small number have been characterized. To establish more generalized understanding their roles in development, we used CRISPR/Cas9 to remove 45 microexons zebrafish assessed larval activity, morphology, behavior. Most mutants had minimal or no phenotypes at this developmental stage. Among previously studied microexons, uncovered baseline stimulus-driven for two (meA meB) ptprd reduced activity telencephalon tenm3 B 0 isoform. mild, neural were discovered several that not characterized, including ppp6r3, sptan1, dop1a, rapgef2, dctn4, vti1a , meaf6 . This study establishes general approach investigating alternative events prioritizes downstream analysis.

Язык: Английский

Процитировано

1
Loss of DOT1L function disrupts neuronal transcription, animal behavior, and leads to a novel neurodevelopmental disorder DOI Creative Commons
Marissa J. Maroni, M. Kathryn Barton,

Katherine A. Lynch

и другие.

medRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Ноя. 2, 2024

ABSTRACT Individuals with monoallelic pathogenic variants in the histone lysine methyltransferase DOT1L display global developmental delay and varying congenital anomalies. However, impact of loss remains unclear. Here, we present a largely female cohort 11 individuals delays dysmorphic facial features. We found that include missense clustered catalytic domain, frameshift, stop-gain variants. demonstrate specific cause activity therefore sought to define effects decreased function. Using RNA-sequencing cultured neurons single nucleus mouse cortical tissue, partial Dot1l depletion causes sex-specific transcriptional responses disrupts transcription synaptic genes. Further, alters neuron branching expression proteins. Lastly using zebrafish models, behavioral disruptions deficits mice. Overall, how leads neurological dysfunction by demonstrating impacts transcription, morphology, behavior across multiple models systems.

Язык: Английский

Процитировано

1
Using zebrafish as an animal model for studying rare neurological disorders: A human genetics perspective DOI Open Access
Dilan Wellalage Don, Tae‐Ik Choi, T. Kim

и другие.

Journal of Genetic Medicine, Год журнала: 2024, Номер 21(1), С. 6 - 13

Опубликована: Июнь 28, 2024

Other SectionsAbstractINTRODUCTIONUNDERSTANDING ZEBRAFISH AS A MODEL ORGANISM IN NEUROSCIENCECANDIDATE GENES AND PHENOTYPICAL ANALYSIS OF DISEASE-RELATED MUTATIONSZEBRAFISH MODELS FOR RARE NEUROLOGICAL DISORDERSCONCLUSIONACKNOWLEDGEMENTSFUNDINGAUTHORS' CONTRIBUTIONSFigureTableReference

Язык: Английский

Процитировано

0
Disrupted development of sensory systems and the cerebellum in a zebrafish ebf3a mutant DOI Creative Commons
Nghi D. P. Dang,

Claire L. Conklin,

Thinh Q. Truong

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Сен. 11, 2024

Abstract Mutations in the transcription factor EBF3 results a neurodevelopmental disorder, and studies animal models indicate that it has critical role neuronal differentiation. The molecular pathways neuron types disrupted by its loss, however, have not been thoroughly investigated. Nor outcomes of these changes on behavior brain activity. Here, we generated characterized zebrafish ebf3a loss-of-function mutant. We discovered morphological neural phenotypes, including an overall smaller size, particularly hypothalamus, cerebellum, hindbrain. Brain function was also compromised, with activity strongly increased cerebellum abnormal at baseline response to visual acoustic stimuli. From RNA-sequencing developing larvae, notable included significant downregulation genes mark olfactory sensory neurons, lateral line, cerebellar Purkinje neurons. This study sets stage for determining which downstream underlie emergence observed phenotypes establishes multiple strong could form basis drug screen.

Язык: Английский

Процитировано

0
Removal of developmentally regulated microexons has a minimal impact on larval zebrafish brain morphology and function DOI Open Access

Caleb CS Calhoun,

Mary E.S. Capps,

Kristie Muya

и другие.

Опубликована: Дек. 24, 2024

Microexon splicing is a vertebrate-conserved process through which small, often in-frame, exons are differentially included during brain development and across neuron types. Although the protein sequences encoded by these highly conserved can mediate interactions, neurobiological functions of only small number have been characterized. To establish more generalized understanding their roles in development, we used CRISPR/Cas9 to remove 45 microexons zebrafish assessed larval activity, morphology, behavior. Most mutants had minimal or no phenotypes at this developmental stage. Among previously studied microexons, uncovered baseline stimulus-driven for two (meA meB) ptprd reduced activity telencephalon tenm3 B 0 isoform. mild, neural were discovered several that not characterized, including ppp6r3, sptan1, dop1a, rapgef2, dctn4, vti1a , meaf6 . This study establishes general approach investigating alternative events prioritizes downstream analysis.

Язык: Английский

Процитировано

0
Removal of developmentally regulated microexons has a minimal impact on larval zebrafish brain morphology and function DOI Open Access

Caleb CS Calhoun,

Mary E.S. Capps,

Kristie Muya

и другие.

Опубликована: Дек. 24, 2024

Microexon splicing is a vertebrate-conserved process through which small, often in-frame, exons are differentially included during brain development and across neuron types. Although the protein sequences encoded by these highly conserved can mediate interactions, neurobiological functions of only small number have been characterized. To establish more generalized understanding their roles in development, we used CRISPR/Cas9 to remove 45 microexons zebrafish assessed larval activity, morphology, behavior. Most mutants had minimal or no phenotypes at this developmental stage. Among previously studied microexons, uncovered baseline stimulus-driven for two (meA meB) ptprd reduced activity telencephalon tenm3 B 0 isoform. mild, neural were discovered several that not characterized, including ppp6r3, sptan1, dop1a, rapgef2, dctn4, vti1a , meaf6 . This study establishes general approach investigating alternative events prioritizes downstream analysis.

Язык: Английский

Процитировано

0