Project description:Bene expression profile of Angus bovine testis tissue at 2, 4 and 8 weeks of age using Affymetrix Bovine GeneChip Experiment Overall Design: Samples obtained from Angus bull calves at 2, 4 and 8 weeks of age. Two replicates at each age, 6 total samples.
Project description:The sex of the animals is of paramount importance in many animal production systems. This is particularly evident in the production of milk or in breeding programs focused on the production of female animals. In some cases, slaughter or euthanasia of animals of the unwanted sex becomes the only solution, highlighting ethical and economic concerns. As global demand for food continues to rise, the importance of addressing these issues becomes more evident. Reproductive technologies, such as sperm sexing techniques, may hold the key to addressing both animal welfare and the sustainability of animal production. The accessibility of cell surface proteins makes them promising targets for innovative sperm sexing techniques. Therefore, a comprehensive shotgun proteomic analysis was performed on protein lysates specifically enriched in cell surface proteins through biotinylation isolation obtained from Limousin bull unsexed semen (BU), Holsteins-Frisian sexed X-semen (BX; purity > 90%), and a pool of Angus and Asturian sexed Y-semen (BY; purity > 90%). According to this analysis, after removal of contaminants, a total of 149 were reliably quantified. BU samples had 18 proteins significantly upregulated and 1 protein (contaminant) significantly downregulated when comparing with BX samples, and 52 proteins significantly upregulated when comparing with BY samples. Regarding possible differences between the X- and Y-sperm proteome, it was found that 5 proteins were significantly upregulated in BX samples. After removal of contaminants, a total of 130 quantified proteins were further analyzed. According to the DeepTMHMM v.1.0.13 software, out of the 130 quantified proteins, 81 were predicted to be globular proteins. Additionally, 23 were predicted as alpha-helical transmembrane proteins, 9 having a signal peptide. A total of 26 proteins were solely predicted to have a signal peptide. Furthermore, through cross-referencing these results with Gene Ontology information obtained from UniProt and one-to-one fast orthology assignments using the eggNOG-mapper v2.1.7 tool, it was possible to identify 28 proteins that could potentially be accessible from the cell surface. Among these are 11 proteins that were identified in BX samples but not in BY samples, making them potential protein targets specific to X-chromosome-bearing spermatozoa. Out of the 11 proteins, 5 have the highest potential for application in sperm sexing techniques as they are predicted to be transmembrane proteins with at least one transmembrane region and were identified in 2 or 3 replicates of the BX samples. Moreover, one of them is significantly more expressed in BX samples than in BY samples. None of the proteins quantified in BY were exclusive of this condition. Moreover, a total of 1,614 deamidated peptides were identified, corresponding to 265 deamidation sites. Of these, 144 deamidated peptides were reliably quantified (3 valid intensity values in one of the experimental conditions), corresponding to 133 unique deamidation sites on 71 proteins. Additionally, 32 possible N-linked glycosylation sites were identified (NxS/T motifs).
Project description:Growth plate chondrocytes were isolated from the distal metacarpus of young dairy cattle (all under 10 mo of age), the chondrocytes were released from the extracellular matrix by digestion with Collagenase P for 4 hours, and the various zones of the growth plate were separated by density centrifugation. The least-dense Hypertrophic Zone (HZ) cells were compared to the most-dense Reserve Zone (RZ) cells. 6 pairs of HZ vs RZ were compared by microarray. Experiment Overall Design: Growth plate chondrocytes were isolated from the distal metacarpus of young dairy cattle (all under 10 mo of age), the chondrocytes were released from the extracellular matrix by digestion with Collagenase P for 4 hours, and the various zones of the growth plate were separated by density centrifugation. The least-dense Hypertrophic Zone (HZ) cells were compared to the most-dense Reserve Zone (RZ) cells. Six independent sample pairs of HZ vs RZ were compared by microarray.
Project description:Wandong cattle are an autochthonous Chinese breed used extensively for beef production. The breed tolerates extreme weather conditions and raw feed and are resistant to tick-borne diseases. However, the genetic basis of testis development and sperm production as well as breeding management is not well established in local cattle. In this study, we performed total RNA-Seq and comprehensively analyzed the circ-RNA expression profiling of the testes samples of six bulls at 3 years and 3 months of developmental age. In total, 17 013 circ-RNAs were identified, of which 681 circRNAs (P-adjust < 0.05) were differentially expressed (DE). Among these DE circ-RNAs, 579 were upregulated and 103 were downregulated in calf and bull testes. The Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses revealed that the identified target genes were classified into three broad functional categories, including biological process, cellular component, and molecular function, and were enriched in the lysine degradation, cell cycle, and cell adhesion molecule pathways.
Project description:Sperm cells are characterized by a unique epigenome, and an incorrect establishment of DNA methylation patterns during the differentiation of male germ cells into spermatozoa has been associated with male infertility in several species. While bull semen is widely used in artificial insemination, the literature describing DNA methylation in bovine sperm is still scarce. The purpose of this study is to compare the methylomes of sperm and somatic cell types in cattle using the RRBS technology.