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Gene expression after deletion of Bmp4 in the mouse optic vesicle

ABSTRACT: Bmp4 was conditionally deleted from the mouse optic vesicle with Rx-Cre. The heads of wild type and knockout E10.5 mouse embryos were laser micodissected to isolate the lens ectoderm and prospective retina from three embryos of each genotype. Total RNA was purified and reverse transcribed and amplified using a NuGEN Ovation Pico WTA system V2 kit. cDNA was biotinylated and hybridized to Illumina Mouse6 V2 bead arrays. Three wild type and three knockout embryos were used. RNA obtained from the lens ectoderm or prospective retina (distal optic vesicle) was pooled, amplified and used for microarray analysis. Statistical analysis was performed using the values from the 25-50 beads on each array.

ORGANISM(S): Mus musculus

SUBMITTER: David Beebe

PROVIDER: E-GEOD-62536 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Gene expression after deletion of Bmp4 in the mouse optic vesicle

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Rat retina and optic nerve LC-MS/MS

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Yap and Taz regulate retinal pigment epithelial cell fate

Project description:The optic vesicle comprises a pool of bi-potential progenitor cells from which the retinal pigment epithelium (RPE) and neural retina fates segregate during ocular morphogenesis. Several transcription factors and signaling pathways have been shown to be important for RPE maintenance and differentiation, but an understanding of the initial fate specification and determination of this ocular cell type is lacking. We show that Yap/Taz-Tead activity is necessary and sufficient for optic vesicle progenitors to adopt RPE identity in zebrafish. A Teadresponsive transgene is expressed within the domain of the optic cup from which RPE arises, and Yap immunoreactivity localizes to the nuclei of prospective RPE cells. yap (yap1) mutants lack a subset of RPE cells and/or exhibit coloboma. Loss of RPE in yap mutants is exacerbated in combination with taz (wwtr1) mutant alleles such that, when Yap and Taz are both absent, optic vesicle progenitor cells completely lose their ability to form RPE. The mechanism of Yap dependent RPE cell type determination is reliant on both nuclear localization of Yap and interaction with a Tead co-factor. In contrast to loss of Yap and Taz, overexpression of either protein within optic vesicle progenitors leads to ectopic pigmentation in a dosagedependent manner. Overall, this study identifies Yap and Taz as key early regulators of RPE genesis and provides a mechanistic framework for understanding the congenital ocular defects of Sveinsson’s chorioretinal atrophy and congenital retinal coloboma. 60 pooled eyes from 36 hpf wild type or vsx2:Gal4/dsRed:14xUAS:YapS87A embryos were pooled for one sample. Three wild type and three vsx2:Gal4/dsRed:14xUAS:YapS87A pools were analyzed for RNA.

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Single-cell transcriptional analysis of lens placode development in Mab21l1 knockout mice

Project description:In mouse early organogenic stages, a Mab21l1-null mutation causes severe defects in invagination of the lens placode toward optic cup due to its severe defects in both cell proliferation and survival, which subsequently results in severe microphthalmia after birth. In human, loss of function variants in MAB21L1 have recently been described in Cerebello-Oculo-Facio-Genital syndrome (COFG syndrome). The close similarity between mouse and human phenotypes including microphthalmia indicates the importance of Mab21l1 function in human lens placode. To approach the molecular function of MAB21L1 in eye morphogenesis, we examined single cell transcriptomes in lens placode and its surrounding surface ectoderm at 26 somite stage (E9.25), just prior to the onset of lens defects in Mab21l1-null embryos. Here we demonstrate the identification of several key genes immediately downstream of Mab21l1 function, together with the early lens placode-/surface ectoderm-specific single cell transcriptomes during murine lens placode specification.

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