mRNA splicing variants of the transcription factor Blimp1 differentially regulate germline genes in echinoderms

Developmental Biology, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 1, 2025

Language: Английский

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ROCK and the actomyosin network control biomineral growth and morphology during sea urchin skeletogenesis

eLife, Journal Year: 2024, Volume and Issue: 12

Published: April 4, 2024

Biomineralization had apparently evolved independently in different phyla, using distinct minerals, organic scaffolds, and gene regulatory networks (GRNs). However, diverse eukaryotes from unicellular organisms, through echinoderms to vertebrates, use the actomyosin network during biomineralization. Specifically, remodeling protein, Rho-associated coiled-coil kinase (ROCK) regulates cell differentiation expression vertebrates’ biomineralizing cells, yet, little is known on ROCK’s role invertebrates’ Here, we reveal that ROCK controls formation, growth, morphology of calcite spicules sea urchin larva. elevated skeletogenic cells downstream Vascular Endothelial Growth Factor (VEGF) signaling. inhibition leads skeletal loss disrupts expression. after spicule formation reduces elongation rate induces ectopic branching. Similar phenotypes are observed when inhibited a culture, indicating these due activity specifically cells. Reduced growth enhanced branching also under direct perturbations network. We propose machinery were employed independently, GRNs, regulate biomineral Eukaryotes.

Language: Английский

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CMTM4 is an adhesion modulator that regulates skeletal patterning and primary mesenchyme cell migration in sea urchin embryos

Developmental Biology, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 1, 2025

Language: Английский

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Single-cell RNA sequencing of the holothurian regenerating intestine reveals the pluripotency of the coelomic epithelium

Published: Feb. 21, 2025

In holothurians, the regenerative process following evisceration involves development of a “rudiment” or “anlage” at injured end mesentery. This regenerating anlage plays pivotal role in formation new intestine. Despite its significance, our understanding molecular characteristics inherent to constituent cells this structure has remained limited. To address gap, we employed state-of-the-art scRNA-seq and HCR-FISH analyses discern distinct cellular populations associated with regeneration anlage. Through approach, successfully identified thirteen cell clusters. Among these, two clusters exhibit consistent putative mesenchymal cells, while another four show features akin coelomocyte populations. The remaining seven collectively form large group encompassing coelomic epithelium Within clusters, recognized previously documented such as muscle precursors, neuroepithelial actively proliferating cells. Strikingly, analysis provides data for identifying least other that define precursor growing Consequently, findings strengthen hypothesis is pluripotent tissue gives rise diverse types intestinal organ. Moreover, results provide initial view into transcriptomic responsible amazing capabilities echinoderms.

Language: Английский

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Single-cell RNA sequencing of the holothurian regenerating intestine reveals the pluripotency of the coelomic epithelium

eLife, Journal Year: 2025, Volume and Issue: 13

Published: March 20, 2025

In holothurians, the regenerative process following evisceration involves development of a ‘rudiment’ or ‘anlage’ at injured end mesentery. This regenerating anlage plays pivotal role in formation new intestine. Despite its significance, our understanding molecular characteristics inherent to constituent cells this structure has remained limited. To address gap, we employed state-of-the-art scRNA-seq and hybridization chain reaction fluorescent situ analyses discern distinct cellular populations associated with regeneration anlage. Through approach, successfully identified 13 cell clusters. Among these, two clusters exhibit consistent putative mesenchymal cells, while another four show features akin coelomocyte populations. The remaining seven collectively form large group encompassing coelomic epithelium Within clusters, recognized previously documented such as muscle precursors, neuroepithelial actively proliferating cells. Strikingly, analysis provides data for identifying least other that define precursor growing Consequently, findings strengthen hypothesis is pluripotent tissue gives rise diverse types intestinal organ. Moreover, results provide initial view into transcriptomic responsible amazing capabilities echinoderms.

Language: Английский

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Automated, high-throughput in-situ hybridization of Lytechinus pictus embryos

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2025, Volume and Issue: unknown

Published: March 24, 2025

Abstract Despite the reach of in situ hybridization (ISH) developmental biology, it has rarely been used at scale. The major limitation throughput assay, which typically relies upon labor intensive manual steps. goal this study was to develop a fully automated chain reaction (HCR) pipeline capable large-scale gene expression pattern profiling, with dramatically reduced cost and effort, sea urchin Lytechinus pictus . Our resulting pipeline, we term high (HT)-HCR, can process 192 probe sets on whole-mount embryos within 32 hours. unique qualities embryo enabled us automate entire HCR assay 96-well plate format, utilize highly miniaturized volumes, general purpose robotic liquid handler, confocal microscopy. From approach produced quality localization data for 101 target genes across three stages L. results reveal previously undescribed physiological genes, as well canonical transcription factors. HT-HCR represents log order increase rate spatial transcriptomic be resolved urchin. This paves way understudied sophisticated perturbation analysis. Summary Statement We developed an high-throughput rapidly map embryos, enabling discovery novel patterns.

Language: Английский

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Ultrafiltration and Fluid Excretion in Echinoids Involves the Axial Organ with Elimination via the Intestine

Life, Journal Year: 2025, Volume and Issue: 15(5), P. 767 - 767

Published: May 10, 2025

Many animals display nephridial structures for the ultrafiltration of metabolic waste. However, a equivalent and an excretory system are not generally recognized echinoderms. Podocytes cells that function in body fluids. Limited ultrastructural analyses echinoderms identify with podocyte morphology axial organ left coelom larval sea urchins. Echinoid internal anatomy suggests functions by organ, as well filtrate flow via water vascular excretion through madreporite; however, these reports based on morphology. To verify podocytes orthologues podocyte-specific genes were evaluated urchin genome RNAseq data sets. from madreporite, fluorescein was used tracer clearance, injected into main cavity Results showed encoding proteins vertebrates expressed specifically urchins, agreement orthologue expression nurse shark kidney. clearance shows elimination anus rather than madreporite. This leads to hypothesis waste occur but flows haemal capillaries intestinal walls, which fluid is transferred lumen anus. Future testing proposed evaluate filtration blastocoel larvae coelom, small or juvenile echinoids have undergone tissue visualize route within cleared, intact

Language: Английский

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Transcription of microRNAs is regulated by developmental signaling pathways and transcription factors

Frontiers in Cell and Developmental Biology, Journal Year: 2024, Volume and Issue: 12

Published: April 24, 2024

In early embryonic development, the cross-regulation of transcription factors and signaling pathways are critical in mediating developmental physiological processes. Additionally, many studies have shown importance post-transcriptional regulation network components mediated by microRNAs (miRNAs); however, how miRNAs transcriptionally regulated is poorly understood. fine-tuners biological processes their dysregulation leads to a variety diseases defects. Previously, we that dynamically expressed throughout sea urchin suggesting likely be under transcriptional regulation. Here, used pharmacological inhibitors, genetic constructs, loss-of-function reagents assess impact key (Wnt, Nodal, MAPK, Sonic Hedgehog, Delta/Notch, VEGF, BMP) (Alx1, Ets1/2, Tbr) on transcript levels evolutionarily conserved miR-1, miR-31, miR-92 miR-124; invertebrate-specific miR-71; echinoderm-specific miR-2002, miR-2007, miR-2012. We also computational methods identify potential factor binding sites these miRNAs. Lists motifs for (TFs) were acquired from MEME-Suite Motif Database as inputs algorithm FIMO (Find Individual Occurrences), which detects short nucleotide within larger sequences. Based experimental data miRNA expression conjunction with bioinformatic predictions, propose Tbr, Alx1, Ets1 regulate SpmiR-1, SpmiR-31, SpmiR-71, respectively. additionally observed significant effects result perturbations Wnt, Hedgehog pathways, while no change or BMP pathways. Overall, this study provides insights into contribute our overall understanding

Language: Английский

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Contrasting the development of larval and adult body plans during the evolution of biphasic lifecycles in sea urchins

Development, Journal Year: 2024, Volume and Issue: 151(20)

Published: Oct. 15, 2024

ABSTRACT Biphasic lifecycles are widespread among animals, but little is known about how the developmental transition between larvae and adults regulated. Sea urchins a unique system for studying this phenomenon because of stark differences their bilateral larval pentaradial adult body plans. Here, we use single-cell RNA sequencing to analyze development Heliocidaris erythrogramma (He), sea urchin species with an accelerated, non-feeding mode development. The time course extends from embryogenesis roughly day before onset metamorphosis in He larvae, which period that has not been covered by previous datasets. We find strategy associated several changes specification cell types compared feeding such as loss larva-specific skeletal population. Furthermore, plans may utilize largely different sets regulatory genes. These findings lay groundwork extending existing gene networks cover additional stages biphasic lifecycles.

Language: Английский

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Structural and molecular distinctions of primary and secondary spines in the sea urchin Lytechinus variegatus

Scientific Reports, Journal Year: 2024, Volume and Issue: 14(1)

Published: Nov. 18, 2024

Sea urchins (echinoids) are common model organisms for research in developmental biology and their unusual transition from a bilaterally organized larva into post-metamorphic adult with pentaradial body symmetry. The also has calcareous endoskeleton multimetameric pattern of continuously added elements, among them the namesake this phylum, spines. Nearly all echinoids have both large primary spines, an associated set smaller secondary We hypothesize that spines tropical variegated urchin species, Lytechinus variegatus, morphologically molecularly distinct structures not just small versions To test premise, we examined spine types using light microscopy, micro-CT imaging, lectin labeling, transcriptomics, fluorescence situ hybridization (FISH). Our findings reveal basic similarities between two mineral cellular anatomy, but clear differences growth patterns, genes expressed, profile various expressed genes. In particular, non-overlapping, longitudinally concentrated bands lead to blunt straight profile, transcriptome involving upregulation many comparison Neural, ciliary, extracellular matrix interacting factors implicated differentially gene (DEG) dataset, including genes—ONECUT2 uncharacterized discoidin- thrombospondin-containing protein. show type-specific localizations by FISH, which will be interest ongoing work patterning. These results demonstrate L. variegatus overlapping molecular biomineralization characteristics, suggesting unique developmental, regenerative, representation spiny dermal phylum.

Language: Английский

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