Integrated analysis of the expression profiles of the lncRNA-miRNA-mRNA ceRNA network in granulosa and cumulus cells from yak ovaries

Abstract

Background Growing oocytes acquire the ability to mature through two-way communication between gametes and surrounding somatic cumulus cells (CCs). Granulosa cells (GCs) support oocyte growth, regulate meiosis progression, and modulate global oocyte transcription activity. However, the proliferation and differentiation of the yak ovary in GCs and CCs remain unclear. To characterize the important roles of long non-coding RNA, (lncRNA), microRNA (miRNA), and messenger RNA (mRNA), whole-transcriptome analysis was performed. Real-time quantitative fluorescence PCR was performed to verify the selected RNA sequences. Results Important gene ontology terms and Kyoto Encyclopedia of Genes and Genomes pathways related to differentiation and oocyte development were identified for the target genes of differentially expressed lncRNAs, miRNAs, and mRNAs. In total,6223 mRNAs (2197 upregulated, 4026 downregulated), 643 lncRNAs (204 upregulated, 479 downregulated), and 559 miRNAs (311 upregulated, 248 downregulated) were significantly altered between the two groups. Target genes involved in cell adhesion, cell differentiation, regulation of developmental processes, cell proliferation, embryo development, signal transduction, apoptosis, and aromatic compound biosynthetic processes were significantly enriched. These RNAs were involved in ECM-receptor interaction, MAPK signaling, Hippo signaling, PI3K-Akt signaling, cell cycle, cell adhesion, leukocyte trans-endothelial migration, and actin cytoskeleton regulation. Conclusions A comprehensive analysis of the co-expression network of competing endogenous RNAs (ceRNAs) will facilitate the understanding of the process of granulosa cell proliferation and differentiation and offer a theoretical basis for the development of oocytes.

Transcriptome Analysis Provides Insights into Copulation, Fertilization, and Gestation in Sebastes schlegelii

“…The potential molecular gene network in oogenesis was conserved among different species with internal fertilization. Previous studies in Atlantic cod [ 27 ], yak [ 28 ], chickens [ 29 ], and insects [ 30 ] also reported similar pathways involved in oogenesis. In addition, in Drosophila [ 31 ], mating-induced oocyte maturation with calcium binding and transport proteins increased, as well as calcium signaling pathways that were also significantly enriched in the present study.…”

Transcriptomics reveals age-related changes in ion transport–related factors in yak lungs

“…The reaction conditions were as follows: 95°C for 5 min, followed by 40 cycles of 95°C for 30 s, 61°C for 30 s, and 72°C for 10 s, and then, by 72°C for 2 min. For each gene, four replicates were analyzed ( 13 ). The relative expression of DEGs was analyzed using the 2 −ΔΔCT method, and the data were analyzed using analysis of variance (ANOVA) on SPSS (version 24.0).…”

“…After they were blocked with skimmed milk at room temperature for 4 h, the membranes were individually incubated with primary antibodies against FOXI1, SLC12A2, and KCNMA1 (each diluted at 1:800) at 4°C overnight. Next, the membranes were washed 10 times with PBST for 5 min, followed by incubation with secondary antibodies (goat antirabbit IgG, diluted at 1:3,000) on a shaker at room temperature for 1.5 h. After another round of 10 washings with PBST for 5 min, the membranes were treated with enhanced chemiluminescence reagent for visualization; next, the images were scanned using a chemiluminescence imager ( 13 ). Band intensity analysis was performed using ImageJ, and the target bands’ grayscale values were compared with those of β-actin bands.…”

Exosomes Derived from Yak Follicular Fluid Increase 2-Hydroxyestradiol Secretion by Activating Autophagy in Cumulus Cells

“…The target and reference primer sequences were designed using Primer Premier 6.0 ( Table 1 ). The relative expression of the target gene transcripts was calculated using the 2 −ΔΔCT method and subjected to statistical analysis using SPSS 22.0 software [ 23 ].…”

“…Then, equal amounts of protein were separated on 10% or 15% SDS-polyacrylamide gel electrophoresis and transferred to polyvinylidene difluoride (PVDF) membranes. The membranes were blocked with 5% skim milk in Tris-buffered saline containing 0.1% Tween-20 for 3 h at room temperature, incubated overnight at 4 °C with primary antibody, and on the next day, they were incubated for 45 min at room temperature with a secondary antibody (Goat Anti-Rabbit IgG-HRP) [ 23 ]. Bands were visualized using an ECL kit (AB65623, Abcam, Cambridge, UK) and detected using the Amersham Imager 600 system (GE Healthcare Life Sciences, Marlborough, MA, USA).…”