Project description:The transcriptome pattern in blastocyst that developed from cumulus oocyte complexes matured in coculture with porcine luteal cells was investigated.
Project description:Comparative trascriptomic analysis between porcine early-blastocyst and Hatched blastocyst collected around day 5-6 direct comparison with dye-swap; two different arrays with 6 Samples each
Project description:This study was designed to investigate the impact of vitrification on the transcriptome profile of blastocysts using a porcine (Sus scrofa) model and a microarray approach. Blastocysts were collected from weaned sows (n = 13). A total of 60 blastocysts were vitrified (treatment group). After warming, vitrified embryos were cultured in vitro for 24 h. Non-vitrified blastocysts (n = 40) were used as controls. After the in vitro culture period, the embryo viability was morphologically assessed. A total of 30 viable embryos per group (three pools of 10 from 4 different donors each) were subjected to gene expression analysis. A fold change cut-off of ±1.5 and a restrictive threshold at p-value < 0.05 were used to distinguish differentially expressed genes (DEGs). The survival rates of vitrified/warmed blastocysts were similar to those of the control (nearly 100%, n.s.). A total of 205 (112 upregulated and 93 downregulated) were identified in the vitrified blastocysts compared to the control group. The vitrification/warming impact was moderate, and it was mainly related to the pathways of cell cycle, cellular senescence, gap junction, and signaling for TFGβ, p53, Fox, and MAPK. In conclusion, vitrification modified the transcriptome of in vivo-derived porcine blastocysts, resulting in minor gene expression changes.
Project description:<p>Melatonin (MT), a neurohormone synthesized and secreted primarily by the pineal gland, is of vital function to animal reproduction. However, the effect of MT treatment on porcine immature Sertoli cells (iSCs) remains unclear. Here, MT treatment (10 nM, 36 h) could elevate mitochondrial function and reduce oxidative stress, therefore inhibit apoptosis of porcine iSCs. Transcriptome profiling identified 39 differentially expressed genes (DEGs) (33 known and 6 novel) (MT vs Control), mainly involved in GO terms of steroid metabolic process, glutamine metabolic process, oxidoreductase activity and G protein coupled receptor binding, and KEGG pathways of steroid biogenesis, pyruvate metabolism, NF-kappa B and AMPK signal pathways. RT-qPCR validated 6 DEGs (Phgdh, Scd, Hmgcs1, Cytb, Pck2 and Sqle) induced by MT to be with the similar change trend to RNA-seq results. The protein level of HMGCS1 and the estradiol level were confirmed to be significantly decreased by MT (10 nM, 36 h) treatment of porcine iSCs. Inhibition of HMGCS1 in porcine iSCs could significantly reduce the level of estradiol. The levels of cholesterol content within cells and lactate in culture medium were unchanged by MT (10 nM, 36 h). 14 metabolites were significantly altered by MT (10 nM, 36 h) treatment of porcine iSCs. Which were involved in multiple vital metabolic pathways. Collectively, MT treatment of porcine iSCs could promote functions of porcine iSCs via modulating gene expression and metabolism.</p>
Project description:<p>Cycloleucine (CL) is a methyl donor inhibitor. Our previous findings showed that CL could affect meiotic maturation and developmental potency of porcine oocytes by reducing nucleic acid N6-methyladenosine (m6A) epigenetic modification level. However, the effects of CL on porcine male reproduction are unclear. Here,we showed that CL treatment of porcine Sertoli cells (SCs) could reduce viability in a dose-dependent manner. CL treatment (40mM, 36h) of porcine SCs also inhibited proliferation, promoted late apoptosis, increased level of intracellular reactive oxygen species (ROS), reduced mitochondrial function, suppressed levels of nucleic acid N6-methyladenosine (m6A), H3K4me3, H3K27ac and H4K16ac, and increased H3K9me2 level. ELISA assays showed that CL (40mM, 36h) decreased lactate production, but unaltered levels of anti-Müllerian hormone and estradiol. RNA-seq and metabolomics identified 1212 differentially expressed genes (DEGs) (536 up- and 676 down-) and 67 significantly different metabolites (SDMs) (45 up- and 22 down-) to be altered by CL (40mM, 36h) treatment of porcine SCs. These DEGs and SDMs were involved in multiple pathways, including PI3K-Akt, cell cycle, Glycolysis/Gluconeogenesis, FoxO, aminoacyl-tRNA biosynthesis etc. Some of DEGs were validated at mRNA and protein levels. Combined analysis of transcriptome and metabolome identified the high correlations between some DEGs and SDMs. These findings indicate that CL could affect functions of porcine SCs by modifying epigenetic modifications to alter gene expression and metabolism.</p>