A stable Netrin-1 fluorescent reporter chicken reveals cell-specific molecular signatures during optic fissure closure.
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ABSTRACT:
PROVIDER: PRJNA1152979 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR30416825_1.fastq.gz | Fastqsanger.gz | |||
| SRR30416825_2.fastq.gz | Fastqsanger.gz | |||
| SRR30416826_1.fastq.gz | Fastqsanger.gz | |||
| SRR30416826_2.fastq.gz | Fastqsanger.gz | |||
| SRR30416827_1.fastq.gz | Fastqsanger.gz |
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Project description:Coloboma is a developmental eye disorder characterised by failure of epithelial fusion during optic fissure closure (OFC). The incomplete identification of genes and molecular pathways that directly regulate fusion restricts current understanding of OFC mechanisms and coloboma causation. The recent identification of pioneer cells at the edges of the fissure margins provides an opportunity to molecularly characterise OFC. Here we describe the generation of a pioneer cell reporter chicken line (NTN1-T2A-eGFP) by targeting green fluorescent protein into the Netrin-1 locus using CRISPR/Cas9 methodology. Our strategy gave 100% transmission of heterozygous (NTN1T2A-eGFP/+) embryos in which GFP localisation faithfully replicated endogenous NTN1 expression. Furthermore, all NTN1T2A-eGFP/+ embryos and hatched birds were phenotypically normal. We then optimised the isolation of GFP expressing cells from embryonic eyes using spectral fluorescence cell-sorting and performed transcriptomic profiling that revealed multiple new pioneer cell markers. Our approach implicated several novel pathways, including HGF/MET signalling, that plausibly mediate fusion in OFC. This work is the first to characterise pioneer cells directly at the molecular level and provides a new chicken reporter line with broad experimental utility.
2025-04-02 | GSE275781 | GEO
Project description:We sought to identify gene expression signatures confined to the small group of cells at the fissure margins that are involved in OFC. Serial cryosections perpendicular to the optic fissure were prepared from mouse embryonic eyes (n=3 at E11.5 and n=3 at E12.5). The fissure margins and a corresponding control region of dorsal optic cup were isolated using laser capture microdissection. This study design aimed to identify the signature of gene expression in the OFC margins and to identify those genes that were more highly expressed along the ventral (inferior) fissure compared to the opposing dorsal (superior region). Fissure closure is active at E11.5 and complete by E12.5
2020-12-17 | E-MTAB-9427 | biostudies-arrayexpress
Project description:Incomplete fusion of the optic fissure leads to ocular coloboma, a congenital eye defect that affects up to 7.5 per 10,000 births and accounts for up to 10 percent of childhood blindness. The molecular and cellular mechanisms that facilitate optic fissure fusion remain elusive. We have profiled global gene expression during optic fissure morphogenesis by transcriptome analysis of tissue dissected from the margins of the zebrafish optic fissure and the opposing dorsal retina before (32 hours post fertilisation, hpf), during (48 hpf) and after (56 hpf) optic fissure fusion. Differential expression analysis between optic fissure and dorsal retinal tissue resulted in the detection of several known and novel developmental genes. The expression of selected genes was validated by qRT-PCR analysis and localisation investigated using in situ hybridisation. We discuss significantly overrepresented functional ontology categories in the context of optic fissure morphogenesis and highlight interesting transcripts from hierarchical clustering for subsequent analysis. We have identified netrin1a (ntn1a) as highly differentially expressed across optic fissure fusion, with a resultant ocular coloboma phenotype following morpholino antisense translation-blocking knockdown and downstream disruption of atoh7 expression. To support the identification of candidate genes in human studies, we have generated an online open-access resource for fast and simple quantitative querying of the gene expression data. Our study represents the first comprehensive analysis of the zebrafish optic fissure transcriptome and provides a valuable resource to facilitate our understanding of the complex aetiology of ocular coloboma.
2020-10-22 | GSE159822 | GEO
Project description:We sought to identify gene expression signatures confined to the small group of cells at the fissure margins that are involved in OFC. Serial cryosections perpendicular to the optic fissure were prepared from human embryonic eyes (n=3, CS17-18, Day41-44(CS 17-18). The fissure margins and a corresponding control region of dorsal optic cup were isolated using laser capture microdissection. This study design aimed to identify the signature of gene expression in the OFC margins and to identify those genes that were more highly expressed along the ventral (inferior) fissure compared to the opposing dorsal (superior region). As CS17-18 (Day41-44) stages have active points of closure they were considered suitable for transcriptomic analyses of the process of human fissure closure.
2020-12-17 | E-MTAB-9426 | biostudies-arrayexpress
Project description:Zfp503/Nlz2 is Required for RPE Differentiation and Optic Fissure Closure
| PRJNA749017 | ENA
Project description:The different stages of the optic fissure can be clearly visualized by making sagittal sections through the mouse eye during early development which represent the optic fissure at open (E10.5), closing (E11.5) and fused (E12.5) states. Laser capture microdissection (LCM) was employed to dissect tissue from the margins of the optic fissure consisting of the outer (presumptive RPE) and inner (presumptive neurosensory retina) layers of the retina. An approximately square-shaped block of optic fissure (50 x 50 mm) was dissected from each side of the fissure. Two rounds of linear amplification were performed on RNA isolated from each of the samples prior to microarray hybridization. Expression data were gathered in biological triplicate at E10.5, E11.5 and duplicate at E12.5, each array representing pooled optic fissure tissue from three embryos from a single litter. Expression signals were ascertained from 45,101 probe sets and normalized across arrays. Keywords: Time course
2008-10-08 | GSE13103 | GEO
Project description:The different stages of the optic fissure can be clearly visualized by making sagittal sections through the mouse eye during early development which represent the optic fissure at open (E10.5), closing (E11.5) and fused (E12.5) states. Laser capture microdissection (LCM) was employed to dissect tissue from the margins of the optic fissure consisting of the outer (presumptive RPE) and inner (presumptive neurosensory retina) layers of the retina. An approximately square-shaped block of optic fissure (50 x 50 mm) was dissected from each side of the fissure. Two rounds of linear amplification were performed on RNA isolated from each of the samples prior to microarray hybridization. Expression data were gathered in biological triplicate at E10.5, E11.5 and duplicate at E12.5, each array representing pooled optic fissure tissue from three embryos from a single litter. Expression signals were ascertained from 45,101 probe sets and normalized across arrays. Experiment Overall Design: Differential gene expression over the course of three days in laser capture microdissected tissue from wild type mouse embryos.
2008-10-26 | E-GEOD-13103 | biostudies-arrayexpress
Project description:Uveal coloboma is a congenital eye malformation caused by failure of the optic fissure to close in early human development. Despite significant progress in identifying genes whose regulation is important for executing this closure, mutations are detected in a minority of cases using known gene panels, implying additional genetic complexity. We show that the mouse Zfp503/Nlz2 gene is dynamically regulated in developing mouse eyes and that loss of its expression results in uveal coloboma. KO embryos exhibit altered expression patterns of several key transcription factors involved in eye development, including Otx2, Mitf, Pax6, Pax2, Vax1 and Vax2, resulting in reduced melanin pigmentation in the presumptive retinal pigment epithelium and its differentiation into a neural-retina-like lineage. From a large cohort of patients with exome sequencing records, only one patient with clinical diagnosis of retinal degeneration was identified carrying a pPresumed loss-of-function variants in the human ortholog, ZNF503. Although no disease-causing association was demonstrated, because of the retinal degeneration rather than of uveal coloboma phenotype, are rare and may be associated with retinal degeneration, rather than a uveal coloboma phenotype, suggesting it is tempting to speculate that a pleiotropic role of ZNF503 might have a pleiotropic role in ocular development and physiology
2022-11-16 | GSE180641 | GEO
Project description:Mice that are mutant for both Fgfr1 and Fgfr2 specifically in the developing retina develop coloboma. To analyze the transcripts that are affected by defective FGF signaling, we micro-dissected the optic fissure region from the control and FGFR condtional mutant mice and did microarray analysis.
2012-10-02 | GSE41260 | GEO
Project description:Mice that are mutant for both Fgfr1 and Fgfr2 specifically in the developing retina develop coloboma. To analyze the transcripts that are affected by defective FGF signaling, we micro-dissected the optic fissure region from the control and FGFR condtional mutant mice and did microarray analysis. E11.5 control and Six3Cre+; Fgfr1fx/fx;Fgfr2 fx/fx optic fissures are dissected using laser-assisted micro-dissection microscope, RNAs are extracted and labelled and hybridyzed to chips
2012-10-02 | E-GEOD-41260 | biostudies-arrayexpress