Project description:With the increasing demand for donkey production, there has been a growing focus on the breeding of donkeys. However, our current understanding of the mechanisms underlying spermatogenesis and maturation in donkeys during reproduction remains limited.In this study, we constructed a single-cell RNA dataset to study the single-cell landscape of donkey spermatogenesis and maturation. This method allows us to analyze the cell composition in testicular and epididymal tissue, providing insights into the changes that occur during donkey spermatogenesis and maturation. In addition, different gene expression signatures associated with various spermatogenic cell types were found
Project description:Donkey milk (DM) has been considered a valuable alternative to human and bovine coun-terparts as well as to infant formulas. Milk extracellular vesicles (EVs) have been proposed to influence key biological processes. The purpose of this study is to provide a compre-hensive characterization of the protein composition of extracellular vesicles (EVs) by ex-tending quantitative proteomic comparisons to EVs derived from donkey colostrum (DC) and mature donkey milk (MDM). The EVs were isolated from DC and MDM samples, characterized, and subjected to proteomic analysis using the tandem mass tag-based quantitative approach
Project description:We investigated the biological effects of ZEA exposure on donkey granulosa cells by using RNA-seq analysis. ZEA at 10 and 30 μM were administered to granulosa cells within 72 hours of in vitro culture. ZEA at 10 μM significantly altered the tumorigenesis associated genes in donkey granulosa cells. Exposure to 10 and 30 μM ZEA treatment significantly reduced mRNA expression of PTEN, TGFβ, ATM, and CDK2 genes, particularly, the ZEA treatment significantly increased the expression of PI3K and AKT genes. Furthermore, immunofluorescence, RT-qPCR, and Western blot analysis verified the gene expression of ZEA-exposed granulosa cells. Collectively, these results demonstrated the deleterious effect of ZEA exposure on the induction of ovarian cancer related genes via the PTEN/PI3K/AKT signaling pathway in donkey granulosa cells in vitro.
Project description:In this study, 3,869 donkey skeletal muscle lncRNAs were identified using RNA-Seq along with a stringent screening procedure in the longissimus dorsi (LD) and gluteal (G) muscles. These lncRNAs share many characteristics with other mammalian lncRNAs, such as shorter open reading frames (ORFs) and lower expression levels than mRNAs. Furthermore, in pairwise comparisons between libraries of the same stage for two genetic types of male Dezhou donkey, 73 differentially expressed lncRNAs were common to all muscle tissues.
Project description:We examined the growth curve, cell cycle, apoptosis and glycolysis of donkey, horse and mule adult fibroblasts (DAFs, HAFs and MAFs), which indicated there are differences in cell proliferation and metabolism. We also derived mule, donkey and horse iPSCs from their respective adult fibroblasts by piggyBac transposition, and we found the induced reprogramming efficiency of mule iPSCs was significantly higher than donkey and horse iPSCs (78.3% vs 58.2% vs 47.9%). miPSCs, diPSCs and hiPSCs all expressed high levels of key endogenous pluripotency genes such as Oct4, Sox2 and Nanog, propagated robustly in single cell passaging and miPSCs were found to proliferated significantly faster than diPSCs and hiPSCs. Furthermore, miPSCs/MAFs clustered closer to diPSCs/DAFs than to hiPSCs/HAFs by RNA-seq. The establishment of miPSCs provide unique experimental materials for further investigation of understanding the “heterosis” and reproductive isolation during speciation.
Project description:Aedes aegypti mosquitoes are efficient vectors of human disease including Zika, Dengue and Chikungunya. After taking a blood meal, proteins from the blood are broken down into amino acids that are essential for egg development and growth. We have discovered a novel role for a neuropeptide receptor in the hindgut that coordinates blood utilization. The hindgut is not a tissue normally implicated in blood digestion, therefore we conducted RNA-seq profiling of the hindgut in wild type females to obtain a transcriptomic profile of these tissues in non-blood fed conditions.
Project description:Digestive system development is orchestrated by combinatorial signaling interactions between endoderm and mesoderm, but how they are integrated in the genome is poorly understood. Here we identified the Xenopus foregut and hindgut progenitor transcriptomes, which are largely conserved with mammals. Using RNA-seq and ChIP-seq we show that BMP/Smad1 regulates dorsal-ventral gene expression in both the endoderm and mesoderm, whereas Wnt/b-catenin acts as a genome-wide toggle between foregut and hindgut programs. In addition to b-catenin-Tcf promoting hindgut gene transcription, we unexpectedly observed Wnt-repressed foregut genes associated with b-catenin-binding to DNA lacking Tcf motifs, suggesting a novel direct repression. We define how BMP and Wnt signaling are integrated in the genome with Smad1 and β-catenin co-occupying DNA elements associated with hundreds of key regulatory genes. These results extend our understanding of GI organogenesis and how Wnt and BMP may coordinate genomic responses in other contexts.
Project description:For definitive endoderm differentiation, iPS cells (day0) were treated with 100 ng ml-1 activin A and 3μM CHIR99021 for 1 day and 100 ng ml-1 activin A for the following two days. Definitive endoderm was subsequently treated with 3uM CHIR99021 and 500ng ml-1 FGF4 for mid/hindgut differentiation. Mid/hindgut floating spheroids (fl; t-Spheroids) were collected from culture medium from day6 to day 8. On day6, mid/hindgut cells were dissociated into single cells and seeded onto EZSPHERE plate to generate suspension spheroids(EZ; s-Spheroids). RNA of fl spheroids and EZ spheroids were extracted using QIAGEN RNeasy kit.