Project description:To test the hypothesis that ablation of the SOX2 in the developing optic cup leads to expansion of peripheral optic cup signaling pathways, we screened whole-genome changes in gene expression in Sox2-mutant eyes compared with controls. Results confirm that ciliary epithelial (peripheral) identity is gained at the expense of neural retinal identity in Sox2-ablated optic cups. Total RNA was isolated from whole eyes enucleated from six wild-type controls and six Sox2-mutant embryos for a total of twelve eyes per genotype. Each embryo (pair of eyes) was analyzed as a single sample.
Project description:To test the hypothesis that ablation of the SOX2 in the developing optic cup leads to expansion of peripheral optic cup signaling pathways, we screened whole-genome changes in gene expression in Sox2-mutant eyes compared with controls. Results confirm that ciliary epithelial (peripheral) identity is gained at the expense of neural retinal identity in Sox2-ablated optic cups.
Project description:Forebrain and optic cups were microdissected from zebrafish embryos at 21 hours post-fertilization (hpf) after heat-shock at 15 hpf to induce overexpression of BMP in heterozygous tg(hsp70l:bmp4) embryos vs wild type controls.
Project description:Murine neutrophils derived from bone marrow of wild-type and cPLA2alpha-knockin mice (with the C1P interaction site of cPLA2alpha ablated) proteomes were compared
Project description:Embryonic stem (ES) cells have a remarkable capacity to self-organize complex, multi-layered optic cups in vitro via a culture technique called SFEBq. During both SFEBq and in vivo optic cup development, Rax (Rx) expressing neural retina epithelial (NRE) tissues utilize Fgf and Wnt/β-catenin signalling pathways to differentiate into neural retina (NR) and retinal-pigmented epithelial (RPE) tissues, respectively. How these signaling pathways affect gene expression during optic tissue formation has remained largely unknown, especially at the transcriptome scale.
Project description:Eye photoreceptor membrane discs in outer rod segments are highly enriched in the visual pigment rhodopsin and the omega-3 fatty acid docosahexaenote (DHA). The eye acquires DHA from blood, but transporters for DHA uptake across the blood-retinal barrier (BRB) or retinal pigment epithelium have not been identified. Mfsd2a is a newly described sodium-dependent lysophosphatidylcholine symporter expressed at the BRB. Microarrays were used to determine difference in gene expression between wild-type and Mfsd2a KO eye cups. RNA was extracted from eye cups from postnatal day 13 wild-type and Mfsd2a KO mice. Equal amounts of RNA from 6 wild-type eye cups were pooled for the wild-type sample, or 6 Mfsd2a KO eye cups were pooled for the Mfsd2a KO sample. Microarray profiling was done on pooled samples with a RIN cut-off of 7.0 using Mouse 430 2.0 arrays (Affymetrix).
Project description:Embryonic stem (ES) cells have a remarkable capacity to self-organize complex, multi-layered optic cups in vitro via a culture technique called SFEBq. During both SFEBq and in vivo optic cup development, Rax (Rx) expressing neural retina epithelial (NRE) tissues utilize Fgf and Wnt/β-catenin signalling pathways to differentiate into neural retina (NR) and retinal-pigmented epithelial (RPE) tissues, respectively. How these signaling pathways affect gene expression during optic tissue formation has remained largely unknown, especially at the transcriptome scale. We generated Day 10 Rx+ optic tissue using SFEBq, exposed these tissues to either Fgf or Wnt/β-catenin stimulation, and assayed their gene expression at Days 12 and 15 using RNA-Seq. We measured gene expression in these 5 sample groups in biological triplicate using RNA-seq (Illumina HiSeq) .