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| Status |
Public on Oct 22, 2020 |
| Title |
Transcriptome profiling of zebrafish optic fissure fusion |
| Organism |
Danio rerio |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
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.
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| Overall design |
We used RNA-seq to identify differentially expressed genes across optic fissure fusion in zebrafish. Samples were isolated retinal regions including the optic fissure (OF) region and control dorsal retina (DR) at timepoints; prefusion (32 hpf), fusing (48 hpf) and post fusion (56 hpf). 5 biological replicates.
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| Contributor(s) |
Owen N, Richardson RE, Toms M, Moosjaee M |
| Citation(s) |
30733552 |
| Submission date |
Oct 21, 2020 |
| Last update date |
Oct 23, 2020 |
| Contact name |
Mariya Moosajee |
| Organization name |
UCL
|
| Department |
Institute of Ophthalmology
|
| Street address |
11-43 Bath Street
|
| City |
London |
| ZIP/Postal code |
EC1V 9EL |
| Country |
United Kingdom |
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| Platforms (1) |
| GPL21741 |
Illumina HiSeq 4000 (Danio rerio) |
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| Samples (30)
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| Relations |
| BioProject |
PRJNA670550 |
| SRA |
SRP288083 |
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