Project description:N6-methyladenosine (m6A) is the most prominent mRNA modification in eukaryotes, and its potential regulatory role has recently been identified in mammals, plants, and yeast. However, how m6A methylation regulates spermatogenesis remains unknown. In this study, cattle-yak testis tissue was used as the experimental material, and the m6A map was generated through preliminary experiments and methylated RNA immunoprecipitation sequencing. Only spermatogonia and Sertoli cells were observed in cattle-yak testis tissue. Experiments examining the expression of methylation-related enzymes and the overall methylation level showed that the methylation level in the testis of the cattle-yak was slightly lower than that of the sexually mature yak, but significantly higher than that of the pre-sexually mature yak. Annotation analysis indicated that differentially methylated peaks were most frequently concentrated in exonic regions, followed by 3'UTR and finally 5'UTR regions. Through enrichment analysis of differentially expressed genes and differentially methylated corresponding genes, GO analysis of T-vs-Y group mainly involved spermatogenesis, including cytoskeleton, actin binding, etc. KEGG analysis showed that the differential genes were mainly enriched in actin cytoskeleton regulation and MAPK signaling pathway. GO analysis of the T-vs-M group mainly involved protein ubiquitination, ubiquitin ligase complexes, ubiquitin-dependent protein catabolism and endocytosis. KEGG analysis mainly involved apoptosis and Fanconi anemia pathways. This study will lay the foundation for elucidating the molecular mechanism of m6A in male sterility of cattle-yak.
Project description:Testis is the most important male reproductive organ, and the integrity of its physiological function is crucial to the successful production of sperm. In this study, the expression profiles of 11 991 and 8 930 cells in testicular tissue of yak and cattle-yak after sexual maturity were established using Single-cell RNA sequencing. The identification results of cell subpopulations and marker genes were analyzed and their possible mechanisms were predicted.
Project description:Cattle-yak is the hybrid offspring of yak and cattle. It has obvious heterosis in production performance, but the male sterility of cattle-yak has always been the focus of attention. Studies have shown that non-coding RNA is involved in the regulation of spermatogenesis. We comprehensively compared the testicular transcription profiles of cattle, yak and cattle-yak. More DEGs, DECs and DEMs were found in the intersection of the two comparison groups of cattle and cattle-yak, yak and cattle-yak, with 4,968, 360 and 59, respectively. The DEGs of cattle-yak, cattle and yak were mainly enriched in biological processes such as spermatogenesis, male gamete generation and sexual reproduction. At the same time, GO and KEGG analysis suggested that DECs host genes and DEMs source genes were also involved in the regulation of spermatogenesis. The construction of potential ceRNA networks found that some differentially expressed ncRNAs may be involved in the regulation of genes related to testicular spermatogenesis, including miR-423-5p, miR-449b, miR-34b/c, miR-15b, etc., as well as unreported miR-6123, miR-1306 and some miRNA and circRNA interaction pairs. This study provides a reference for further study on the mechanism of male sterility in cattle-yak.
Project description:This study used yak and cattle-yak testes from different developmental stages as materials to construct a complete translation map of the testes, and integrated transcriptome and translation results to explore gene expression changes during the sexual maturation process of yak testes. This study utilized Ribo seq technology to construct a transcriptome map of yak testicular development, revealing that the expression of genes related to spermatogenesis is specifically translated and regulated at different developmental stages. In addition, many unknown open reading frames (ORFs) in the testes have been newly identified.
Project description:Studying spermatogenesis is the key to understanding the development mechanism of the yak reproductive system. Although N6-methyladenosine (m6A) RNA modification has been reported to regulate spermatogenesis and reproductive function in mammals, the molecular mechanism of m6A in yak testis development and spermatogenesis remains largely unknown. Therefore, we collected testicular tissue from junile and adult yaks, and found that the m6A level significantly increased after sexual maturity in yak. In MeRIP-seq, 1,702 hypermethylated peaks and 724 hypomethylated peaks were identified. The hypermethylated differentially methylated RNAs (DMRs) (CIB2, AK1, FOXJ2, PKDREJ, SLC9A3, and TOPAZ1) mainly regulated spermatogenesis, while the hypomethylated DMRs (BCL6, USP25, CD96, EPHA2, and TAF12) mainly participated in immune response. Functional enrichment analysis showed that DMRs were significantly enriched in the adherens junction, gap junction, and Wnt, PI3K, and mTOR signaling pathways, regulating cell development, spermatogenesis, and testicular endocrine function. The functional analysis of differentially expressed genes showed that they were involved in the biological processes of mitosis, meiosis, and flagellated sperm motility during sexual maturity of yak testis. We also screened the key regulatory factors of testis development and spermatogenesis by combined analysis, which included BRCA1, CREBBP, STAT3, and SMAD4. This study has important research value and practical guiding significance for yak genetic improvement, particularly in understanding the molecular mechanism and evolutionary dynamics of the progression of testis developmental stages in yaks.
Project description:This study explores how yaks, an ideal animal model for studying plateau adaptability, adapt to high-altitude environments. The lung is a representative organ of the yak’s adaptation to high-altitude environments. The F1 hybrids of yak and cattle, known as dzho, also exhibit adaptability to plateau conditions. This study constructed a single-cell transcriptome atlas of the lungs in yak, dzho and cattle, containing 51 subtypes. We initially found that the differential subtypes among yak, dzho and cattle were mainly concentrated in T&NK cells and fibroblasts. Most of them belonged to new cell subtypes. Subsequently, we speculated that NKG7 and CD4 played a central regulatory role in T&NK cells, contributing to T cell activation and affecting the adaptive immune response of yaks. In fibroblasts, MYL9 and IGF2 are believed to play key regulatory roles, participating in maintaining cellular energy metabolism balance. These results provide an important scientific basis for systematically analysing the molecular regulatory mechanisms of yak adaptation to high-altitude hypoxia.
Project description:Purpose:Yak long-term colonization and widespread distribution across the plateau can be serve as an ideal natural animal model to provide insights into the adaptive evolution of other plateau species, including humans. Methods:To exploring the molecular mechanisms of lung tissue in yak to response to hypoxia, the mRNA, lncRNA and miRNA of lung tissue from cattle and three different altitude yaks were sequenced. Results:A total of 21764 mRNAs, 14168 lncRNAs and 1209miRNAs (305 known and 904 novel miRNAs)were identifed.Compared yak with cattle, 4975 mRNAs, 3326 lncRNAs and 75 miRNAs were differentially expressed. 756 mRNAs, 346 lncRNAs and 83 miRNAs were found to be differentially expressed amongthree different altitude yaks(fold change≥2 and P-value<0.05). Conclusions:The differentially expressed genes were functionally enriched in long-chain fatty acid metabolic process and protein processing between yak and cattle, while the immune response and cell cycle were enriched among three different altitude yaks. Furthermore, the competing endogenous RNAs (ceRNAs) networks were identified to illustrate their roles.
Project description:Sex condition has been demonstrated to alter meat quality and sex is a major factor that affects the fatty acid composition of lipids of carcass dissectible or intramuscular depot fats. But the possible genetic molecular mechanism of gender causing meat quality differences is not well defined. Qinchuan cattle, Qinghai yak and Guangxi buffalo are three typical indigenous species of cattle in China. Obivious differences of meat quality exist among the three species of cattle. Few studies have been conducted to elucidate the muscle tissue expression of genes involved in pathways and mechanisms leading to meat quality differences beyond the phenotype properties of beef. Bovine Genome Arrays were used to construct muscle expression profiles of the longuissimus dorsi from Qinchuan cattle at 36 months and screen differentially expressed genes in the longuissimus dorsi muscle tissues among different genders of Qinchuan cattle, between Qinchuan cattle and Qinghai yak, and between Qinchuan cattle and Guangxi buffalo.
Project description:Purpose: The goal of this study was to reveal epigenetic differences in the microRNA transcriptomes of two organs (heart and lung) between yak and cattle. Methods: Three unrelated 2-year old adult females for both of yaks and cattle (Luxi Huang cattle) were used in this study. Two of significant hypoxia-responsive tissues (heart and lung) were rapidly collected from each carcass, washed three times with physiological saline, immediately frozen in liquid nitrogen. All frozen samples were stored at –80 °C until RNA extraction.The total RNA were extracted with Trizol (Ambion, USA). NanoDrop ND-2000 spectrophotometer (Nano Drop, DE, USA) and Bioanalyzer 2100 (Agilent Technologies, Santa Clara, CA, USA) were used to monitor the concentration and integrity of RNA, respectively. In brief, several successive steps consist the Illumina sequencing. The small RNA with length of 14-40 nt were first purified by polyacrylamide gel electrophoresis (PAGE), and then specific adapters were ligated to the purified small RNA. The ligated RNA were reverse transcribed to cDNA libraries. Finally, each library were sequenced on Genome Analyzer. Results: We identified 808 widely-expressed conserved and 697 species-specific novel miRNAs in two species. In addition, although two organs showed similar high expression miRNAs, larger differentiation was present in lung than heart between two species. In addition, miRNAs with significantly differentiated patterns of expression in two organs exhibited obvious co-operation effect in high altitude adaptation in form of miRNA family and cluster. Functional analysis revealed that a large amount of differentially expressed miRNAs were enriched in hypoxia-related pathways, such as VEGF signaling pathway, HIF-1α signaling pathway, insulin signaling pathway, DNA damage response, apoptosis, fatty acid metabolism and glucose metabolism. These results suggested the diverse degrees of epigenetic variation in different tissues between yak and cattle, and revealed extensive roles of miRNAs in high altitude adaptation. Conclusions: In this study, we illustrated the differences in the microRNA transcriptomes level for heart and lung between yak and cattle, and suggested extensive roles of miRNAs in high altitude adaptation. The work performed here will provide a typical demonstration for future deciphering the mechanism of high altitude adaptation
Project description:Purpose:Yak long-term colonization and widespread distribution across the plateau can be serve as an ideal natural animal model to provide insights into the adaptive evolution of other plateau species, including humans. Methods:To exploring the molecular mechanisms of lung tissue in yak to response to hypoxia, the mRNA, lncRNA and miRNA of lung tissue from cattle and three different altitude yaks were sequenced. Results:A total of 21764 mRNAs, 14168 lncRNAs and 1209miRNAs (305 known and 904 novel miRNAs)were identifed.Compared yak with cattle, 4975 mRNAs, 3326 lncRNAs and 75 miRNAs were differentially expressed. 756 mRNAs, 346 lncRNAs and 83 miRNAs were found to be differentially expressed amongthree different altitude yaks(fold change≥2 and P-value<0.05). Conclusions:The differentially expressed genes were functionally enriched in long-chain fatty acid metabolic process and protein processing between yak and cattle, while the immune response and cell cycle were enriched among three different altitude yaks. Furthermore, the competing endogenous RNAs (ceRNAs) networks were identified to illustrate their roles.