Project description:Deep sequencing of mRNA from 6 organs of yak (Bos grunniens) Analysis of ploy(A)+ RNA of brain,heart,liver,lung,spleen, and stomach of yak (Bos grunniens)
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:The Gayal (Bos frontalis) is a rare semi-domesticated cattle in China. Gayal has typical beef body shape and good meat production performance. Compared with other cattle species, it has the characteristics of tender meat and extremely low fat content. To explore the underlying mechanism responsible for the differences of meat quality between different breeds, the longissimus dorsi muscle (LM) from Gayal and Banna cattle (Bos taurus) were investigated using transcriptome analysis. The gene expression profiling identified 638 differentially expressed genes (DEGs) between LM muscles from Gayal and Banna cattle. Gene Ontology (GO) enrichment of biological functions and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that the gene products were mainly involved in the PPAR signaling pathway, lipid metabolism and amino acid metabolism pathway. Protein-protein interaction(PPI) network analysis showed APOB, CYP7A1, THBS2, ITGAV, IGFBP1 and IGF2R may have great impact on meat quality characteristics of Gayal. Moreover, three transcription factors, FOXA2, NEUROG2, and RUNX1, which may affect meat quality by regulating the expression of genes related to muscle growth and development have also been found. In summary, our research reveals the molecular mechanisms that cause Gayal meat quality characteristics. It will contribute to improving meat quality of cattle through molecular breeding.
Project description:Alternative splicing (AS) is strictly regulated during cell differentiation and development. AS events are common in the testis, but the splicing regulation in spermatogenesis is still unclear. In this experiment, the third-generation ONT sequencing was used to sequence the full-length transcriptome of testicular tissue, and 40,038 new transcripts were obtained, and the proportion was almost close to the number of known transcripts identified. A total of 7,645 fused transcripts, 15,355 ASs, 25,798 SSRs, and 35,503 lncRNAs were detected. Through gene co-expression network analysis, the key pathways and Hub genes in each stage of yak testicular development were confirmed. The effects of alternative splicing and splicing variation on mammalian spermatogenesis will provide new insights into the potential application of alternative splicing in the treatment of male infertility.