Polarity and morphogenesis of the eye epithelium requires the adhesion junction associated adaptor protein Traf4

Hehr, Carrie L.; Halabi, Rami; McFarlane, Sarah

doi:10.1080/19336918.2018.1477900

Cited by 4 publications

(5 citation statements)

References 52 publications

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“…We have mapped the integration sites in Tg(myl7:Lifeact-GFP) mb22 and Tg(myl7:Lifeact-GFP) mb23 to the cadherin2 and traf4a genes, respectively. We do not think the heart phenotype in these lines was caused by disruption of these host genes at the integration sites because the heterozygous mutant of cadherin2 or traf4a in zebrafish showed no edema phenotypes ( Brusés, 2011 ; Hehr et al, 2018 ), and moreover, the edema phenotype in Tg(myl7:Lifeact-GFP) mb22 could be rescued by knockdown of Lifeact-GFP expression.…”

Section: Discussionmentioning

confidence: 99%

“…In the Tg(myl7:Lifeact-GFP) mb23 transgenic line, the transgene was integrated into the last exon of traf4a (Figure 4B), which encodes tumor necrosis factor (TNF)-receptor-associated factor-4 (TRAF4). Previous studies showed that Cadherin2 or traf4a heterozygous mutants showed normal development and could survive to adult stage without any cardiac abnormality (Brusés, 2011;Hehr et al, 2018), indicating that the cardiac muscle defects observed in Tg(myl7:Lifeact-GFP) mb22 and Tg(myl7:Lifeact-GFP) mb23 hemizygous transgenic lines were unlikely caused by the transgene integration in these genes.…”

Section: Characterization Of the Tg(myl7:lifeact-gfp) Integration Sites In Transgenic Lines With Heart Defectmentioning

confidence: 95%

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Overexpression of Lifeact-GFP Disrupts F-Actin Organization in Cardiomyocytes and Impairs Cardiac Function

2021

Front. Cell Dev. Biol.

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Lifeact-GFP is a frequently used molecular probe to study F-actin structure and dynamic assembly in living cells. In this study, we generated transgenic zebrafish models expressing Lifeact-GFP specifically in cardiac muscles to investigate the effect of Lifeact-GFP on heart development and its application to study cardiomyopathy. The data showed that transgenic zebrafish with low to moderate levels of Lifeact-GFP expression could be used as a good model to study contractile dynamics of actin filaments in cardiac muscles in vivo. Using this model, we demonstrated that loss of Smyd1b, a lysine methyltransferase, disrupted F-actin filament organization in cardiomyocytes of zebrafish embryos. Our studies, however, also demonstrated that strong Lifeact-GFP expression in cardiomyocytes was detrimental to actin filament organization in cardiomyocytes that led to pericardial edema and early embryonic lethality of zebrafish embryos. Collectively, these data suggest that although Lifeact-GFP is a good probe for visualizing F-actin dynamics, transgenic models need to be carefully evaluated to avoid artifacts induced by Lifeact-GFP overexpression.

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Section: Discussionmentioning

confidence: 99%

Section: Characterization Of the Tg(myl7:lifeact-gfp) Integration Sites In Transgenic Lines With Heart Defectmentioning

confidence: 95%

Overexpression of Lifeact-GFP Disrupts F-Actin Organization in Cardiomyocytes and Impairs Cardiac Function

2021

Front. Cell Dev. Biol.

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“…The foxa2 probe was a gift from Dr. Peng Huang (University of Calgary). The fgf8a , pax6a , foxg1a and sema3fa probes were made as described previously 8,30,31 …”

Section: Methodsmentioning

confidence: 99%

“…The fgf8a, pax6a, foxg1a and sema3fa probes were made as described previously. 8,30,31 The in situ hybridization protocol was adapted from existing methods. 32 Minor modifications included incubating the embryos in anti-DIG-AP Fab fragments (Roche) in blocking buffer at room temperature (RT) for 2 h and then leaving the embryos in phosphate buffered saline/0.1% tween (PBST) overnight at 4 C. Embryos were washed in alkaline buffer, transferred to 24-well culture dishes (Thermo Scientific), and stained with NBT/BCIP (Roche) in a 4.5/3.5 μl/ml ratio, respectively.…”

Section: Mrna In Situ Hybridizationmentioning

confidence: 99%

Plexina4 and cell survival in the developing zebrafish hindbrain

Nurcombe

Hehr

McFarlane

2023

Developmental Dynamics

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BackgroundGrowth factors are important in the developing and mature nervous system to support the survival of neurons. Developmental signaling molecules are known for their roles in controlling neurogenesis and neural circuit formation. Whether or not these molecules also have roles in cell survival in the developing nervous system is poorly understood. Plexins are a family of transmembrane receptors that bind Semaphorin ligands and are known to function in the guidance of developing axons and blood vessels.ResultsIn embryonic zebrafish, plexina4 is expressed widely in the brain, becoming largely restricted to the hindbrain as neurogenesis and differentiation proceed. Apoptosis is increased in the embryonic hindbrain of a plexina4ca307/ca307 CRISPR mutant. Based on the literature, we tested the secreted heat shock protein, Clusterin, as a candidate ligand to mediate cell survival through Plexina4. clusterin is expressed by the floor plate of the embryonic zebrafish hindbrain, in proximity to plexina4‐expressing hindbrain cells. Morpholino‐mediated knockdown of Clusterin increases cell apoptosis in the hindbrain, with additional cell death observed in epistasis experiments where Clusterin is knocked down in a plexina4 mutant background.ConclusionsOur data suggest that Plexina4 promotes cell survival in the developing zebrafish hindbrain, likely through a pathway independent of Clusterin.

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“…In some extreme cases, the injected embryos exhibited incomplete dual axes (arrows in Fig 7A). We further examined expression of the genes that mark dorsally derived tissues, including pax2.1 (marking midbrain-hindbrain boundary), krox20 (labeling rhombomeres 3 and 5), myoD (dorsal mesoderm), and six3a (forebrain) [37][38][39][40]. We observed clear lateral expansion of the midbrain-hindbrain boundary and rhombomeres 3 and 5 in the zfp655 mRNA-injected zebrafish embryos at the 10 somite stage (Fig 7C).…”

Section: Overexpression Of Amphioxus Zfp665 and Soxb1b Dorsalizes Zebmentioning

confidence: 96%

Molecular asymmetry in the cephalochordate embryo revealed by single-blastomere transcriptome profiling

Lin

et al. 2020

PLoS Genet

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Studies in various animals have shown that asymmetrically localized maternal transcripts play important roles in axial patterning and cell fate specification in early embryos. However, comprehensive analyses of the maternal transcriptomes with spatial information are scarce and limited to a handful of model organisms. In cephalochordates (amphioxus), an early branching chordate group, maternal transcripts of germline determinants form a compact granule that is inherited by a single blastomere during cleavage stages. Further blastomere separation experiments suggest that other transcripts associated with the granule are likely responsible for organizing the posterior structure in amphioxus; however, the identities of these determinants remain unknown. In this study, we used high-throughput RNA sequencing of separated blastomeres to examine asymmetrically localized transcripts in two-cell and eight-cell stage embryos of the amphioxus Branchiostoma floridae. We identified 111 and 391 differentially enriched transcripts at the 2-cell stage and the 8-cell stage, respectively, and used in situ hybridization to validate the spatial distribution patterns for a subset of these transcripts. The identified transcripts could be categorized into two major groups: (1) vegetal tier/germ granule-enriched and (2) animal tier/anterior-enriched transcripts. Using zebrafish as a surrogate model system, we showed that overexpression of one animal tier/anterior-localized amphioxus transcript, zfp665, causes a dorsalization/anteriorization phenotype in zebrafish embryos by downregulating the expression of the ventral gene, eve1, suggesting a potential function of zfp665 in early axial patterning. Our results provide a global transcriptomic blueprint for early-stage amphioxus embryos. This dataset represents a rich platform to guide future characterization of molecular players in early amphioxus development and to elucidate conservation and divergence of developmental programs during chordate evolution.

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Polarity and morphogenesis of the eye epithelium requires the adhesion junction associated adaptor protein Traf4

Cited by 4 publications

References 52 publications

Overexpression of Lifeact-GFP Disrupts F-Actin Organization in Cardiomyocytes and Impairs Cardiac Function

Overexpression of Lifeact-GFP Disrupts F-Actin Organization in Cardiomyocytes and Impairs Cardiac Function

Plexina4 and cell survival in the developing zebrafish hindbrain

Molecular asymmetry in the cephalochordate embryo revealed by single-blastomere transcriptome profiling

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