Project description:High-throughput sequencing of small RNAs in chicken somite embryos Dissected tissue white leghorn chicken embryos was disaggregated and RNA extracted using the miRVana kit (Ambion). Small RNA fraction between 19 and 24 nt was isolated from 15% denaturing polyacrylamide gel and 15 micro gram was ligated to Solexa adaptors (Illumina) without de-phosphorylating and re-phosphorylating. The short RNAs were converted to DNA by RT-PCR following the Illumina protocol.
Project description:We applied Illumina sequencing to identify microRNAs (miRNAs) and piwi-interacting small RNAs (piRNAs) in the pre- and post-gastrula chicken embryos, and simultaneously obtain a comprehensive expression profile of miRNAs at diverse developmental stages.
Project description:We applied Illumina sequencing to identify microRNAs (miRNAs) and piwi-interacting small RNAs (piRNAs) in the pre- and post-gastrula chicken embryos, and simultaneously obtain a comprehensive expression profile of miRNAs at diverse developmental stages. Discovery and characterization of small RNA species from pre-gastrula and post-gastrula chicken embryos through deep sequencing
Project description:Capture-C from in situ HiC libraries in tailbud tissue containing pre-somitic mesoderm (PSM) dissected from Smchd1GFP/GFP, Smchd1MD43-GFP/MD43-GFP, Smchd1+/+ and Smchd1MD43/MD43 embryos E8.5 embryos at the 7-9 somite stage of development. Somite stage is a good indicator of the precise stage of embryonic development.
Project description:Integrins are a major class of heterodimeric adhesion receptors composed of an a and b subunit that mediate cell adhesion to the extracellular matrix (ECM). The extracellular matrix protein fibronectin is important for early vertebrate development, and Integrin a5b1 and aVb3 are the two primary fibronectin receptors. To better define the integrin – ECM protein network at 10-13 somite stage of zebrafish development, we performed co-immunoprecipitation and Mass Spectrometry (MS) based proteomics using FLAG-tagged Integrin a5, aV, and aVb3 expressed in maternal zygotic a5 mutant (MZa5-/-) embryos. We found that Integrin a5b1 and aVb1 are the functional fibronectin receptors, whereas Integrin aVb3 displayed low affinity to both fibronectins (Fn1a and Fn1b). In addition, basement membrane ligands Laminins (lama1, lamb1a, lamc1) are roughly equal in all three datasets while Thrombospondins (thbs3b, thbs4b) and cartilage oligomeric matrix protein (comp/thbs5) are found exclusively in the aV dataset. Our results suggest a diverse role of aV class integrins in ECM protein recruitment.
Project description:In order to profile periderm-specific nucleosome free regions of zebrafish periderm cells at 4-somite stage, we isolated the equal number of GFP-positive cells and GFP-negative cells from Tg(krt4:GFP) embryos and perform ATAC-seq. This study revealed the landscape of zebrafish periderm-specific nucleosome free regions at 4-somite stage.
Project description:In this study, methylated DNA immunoprecipitation and high-throughput sequencing (MeDIP-seq) was used to provide an atlas of DNA methylomes in the heart tissue of hypoxic highland Tibetan and lowland Chahua chicken embryos.A total of 31.2 gigabases (Gb) of sequence data were generated from six MeDIP-seq libraries. We identified 1049 differentially methylated regions (DMRs) and 695 related differentially methylated genes (DMGs) between the two chicken breeds. The DMGs were involved in vascular smooth muscle contraction, VEGF signaling pathway, calcium signaling pathway, and other hypoxia related pathways. Five candidate genes that had low methylation (EDNRA, EDNRB, BMPR1B, BMPRII, and ITGA2) might have key regulatory functions for hypoxia adaptation in Tibetan chicken embryos. Our study provides significant explanations for the functions of genes and their epigenetic regulation for hypoxic adaptation in Tibetan chickens.