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Gene expression effects of morpholino directed spRunt-1 knockdown on hatched blastula stage embryos

ABSTRACT: Runt domain (Runx) transcription factors constitute a metazoan family of sequence specific DNA binding proteins that are essential for animal development. The sea urchin Runx gene SpRunt-1, which is expressed globally during early embryogenesis, is required for blastula stage cell proliferation and subsequently for cell survival. To obtain a comprehensive list of SpRunt-1 regulatory targets we screened a custom microarray representing the blastula stage transcriptome of Strongylocentrotus purpuratus, comparing relative hybridization signals generated by labeled RNA from 18 hr control versus SpRunt-1 morphant embryos.

ORGANISM(S): Strongylocentrotus purpuratus

PROVIDER: GSE19751 | GEO | 2010/01/07

SECONDARY ACCESSION(S): PRJNA122219

REPOSITORIES: GEO

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Gene expression effects of morpholino directed spRunt-1 knockdown on hatched blastula stage embryos

Project description:Runt domain (Runx) transcription factors constitute a metazoan family of sequence specific DNA binding proteins that are essential for animal development. The sea urchin Runx gene SpRunt-1, which is expressed globally during early embryogenesis, is required for blastula stage cell proliferation and subsequently for cell survival. To obtain a comprehensive list of SpRunt-1 regulatory targets we screened a custom microarray representing the blastula stage transcriptome of Strongylocentrotus purpuratus, comparing relative hybridization signals generated by labeled RNA from 18 hr control versus SpRunt-1 morphant embryos. For an 8x15k chip the 8 arrays were 4 biological replicates and a dye swap of the same 4. Biological Replicates - Four separate embryo cultures, each from a different mating pair were initiated and divided into a control group and a treatment group. All of the embryos in the treatment group were micoinjected with a splice-blocking morpholino antisense oligonucleotide (MASO) targeted to SpRunt-1. All control and sample embryos were developed to hatched blastula stage (18 hours post-fertilization) in filtered seawater at 15Â°C. Total RNA was extracted from each of the four separate samples (3,600 embryos in each sample) with an RNA yield near 5 micro grams per sample. Total RNA was also extracted from the untreated embryos of each separate culture pooled together (about 14,400) with an RNA yield near 20 micro grams. Technical (dye swap) Replicates - Control derived Cy3 (green) labeled aRNA was mixed with sample derived Cy5 (red) labeled aRNA in one biological replicate set of 4 samples and using half of the extracted RNA amount. The other half of the RNA was reciprocally labeled with control derived aRNA Cy5 labeled and sample derived aRNA Cy3 labeled.

2010-05-16 | E-GEOD-19751 | biostudies-arrayexpress

Response to elevated pCO2 in Strongylocentrotus purpuratus larvae

2013-01-01 | GSE37522 | GEO

Synergistic effects of temperature and pCO2 on growth, development and gene expression of Strongylocentrotus purpuratus larvae

Project description:Transcriptional profiling of purple sea urchin (Strongylocentrotus purpuratus) larvae cultured under four different seawater conditions: (i)13°C/400 µatm pCO2, (ii)13°C/1100 µatm pCO2, (iii)18°C/400 µatm pCO2 (iv)18°C/1100 µatm pCO2. The goal was to determine the effects of temperature and CO2, both important climate change variables, on gene expression

2013-01-01 | GSE39125 | GEO

Response to elevated pCO2 in Strongylocentrotus purpuratus larvae

Project description:Transcriptional profiling of purple sea urchin (Strongylocentrotus purpuratus) larvae cultured under three different seawater CO2 concentrations 400, 800, 1200 M-BM-5atm. The goal was to determine the effects of CO2, an important climate change variable, on global gene expression Larvae were cultured under three different seawater CO2 concentrations 400, 800, 1200 M-BM-5atm, each with four replicate cultures, and sampled at two developmental stages (gastrula and pluteus)

2013-01-01 | E-GEOD-37522 | biostudies-arrayexpress

Global patterns of enhancer activity during sea urchin embryogenesis assessed by eRNA profiling

Project description:We used CAGE-seq (Capped Analysis of Gene Expression with Sequencing) to profile eRNA expression and enhancer activity during embryogenesis of the sea urchin, Strongylocentrotus purpuratus. We identified >18,000 enhancers that were active during late oogenesis and early development and documented a burst of enhancer activation during cleavage and early blastula stages. Most enhancers were located near gene bodies and eRNA expression levels were highest for elements near core promoters. Transcriptional signals from enhancers generally paralleled the expression levels of likely target genes. Furthermore, enhancers near lineage-specific genes contained signatures of inputs from developmental gene regulatory networks deployed in those lineages. A large fraction (60%) of sea urchin enhancers previously shown to be active in transgenic reporter assays were associated with eRNA expression. Moreover, a large fraction (50%) of a representative subset of enhancers identified by eRNA profiling drove tissue-specific gene expression in isolation when tested by reporter assays. Our findings provide an atlas of developmental enhancers in a model sea urchin and support the utility of eRNA profiling as a tool for enhancer discovery and regulatory biology. The data generated in this study are publicly available at Echinobase (www.echinobase.org).

2021-06-30 | GSE169227 | GEO

Select microRNAs are essential for early development in the sea urchin.

Project description:microRNAs (miRNAs) are small noncoding RNAs that mediate post-transcriptional gene regulation and have emerged as essential regulators of many developmental events. The transcriptional network during early embryogenesis of the purple sea urchin, Strongylocentrotus purpuratus, is well described and would serve as an excellent model to test functional contributions of miRNAs in embryogenesis. We examined the loss of function phenotypes of the major components of the miRNA biogenesis pathway. Inhibition of de novo synthesis of Drosha and Dicer in the embryo led to consistent developmental defects, a failure to gastrulate, and embryonic lethality, including changes in the steady state levels of transcription factors and signaling molecules involved in germ layer specification. We annotated and profiled small RNA expression from the ovary and several early embryonic stages by deep sequencing followed by computational analysis. All miRNAs have dynamic accumulation profiles through early development as do a large population of putative piRNAs (piwi-interacting RNAs). Defects in morphogenesis caused by loss of Drosha can be rescued with four miRNAs which permits a strong miRNA functional assay. Taken together our results indicate that post-transcriptional gene regulation directed by miRNAs is functionally important for early embryogenesis and is an integral part of the early embryonic gene regulatory network in S. purpuratus. Small RNA expression profiling in early sea urchin development

2012-02-17 | E-GEOD-35876 | biostudies-arrayexpress

Synergistic effects of temperature and pCO2 on growth, development and gene expression of Strongylocentrotus purpuratus larvae

Project description:Transcriptional profiling of purple sea urchin (Strongylocentrotus purpuratus) larvae cultured under four different seawater conditions: (i)13M-BM-0C/400 M-BM-5atm pCO2, (ii)13M-BM-0C/1100 M-BM-5atm pCO2, (iii)18M-BM-0C/400 M-BM-5atm pCO2 (iv)18M-BM-0C/1100 M-BM-5atm pCO2. The goal was to determine the effects of temperature and CO2, both important climate change variables, on gene expression Larvae were cultured under four different seawater conditions (ii)13M-BM-0C/1100 M-BM-5atm pCO2, (iii)18M-BM-0C/400 M-BM-5atm pCO2 (iv)18M-BM-0C/1100 M-BM-5atm pCO2, (i)13M-BM-0C/400 M-BM-5atm each with three replicate cultures for each condition and sampled at pluteus stage

2013-01-01 | E-GEOD-39125 | biostudies-arrayexpress

Base-resolution DNA methylation maps of purple sea urchin (Strongylocentrotus purpuratus)

Project description:Base-resolution DNA methylation maps of purple sea urchin (Strongylocentrotus purpuratus)

| PRJNA778394 | ENA

Strongylocentrotus purpuratus

Project description:Strongylocentrotus purpuratus RNAseq

| PRJNA322662 | ENA

An ancestral Wnt-Brachyury feedback loop and vertebrate-specific recruitment of mesoderm-determining target genes revealed by comparative Brachyury target screens

Project description:Transcription factor brachyury plays a key role during mesoderm development in vertebrates. In this study we study study the effect of this transcription factor in the purple sea urchin S. purpuratus.

2022-08-01 | GSE198320 | GEO

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