Comparative iTRAQ proteomics identified proteins associated with sperm maturation between yak and cattleyak epididymis.

BACKGROUND: During maturation, spermatozoa acquire motility and fertilizing capacity as they transit through the epididymis. In recent years, two-dimensional gel electrophoresis has been employed in proteomics studies conducted in rat, boar and human. However, there has not been a complete information regarding the proteins associated with sperm maturation in the epididymis. In this study, we employed iTRAQ proteomics to investigate proteins associated with sperm maturation between yak and cattleyak epididymis. RESULTS: After a successful sampling and protein extraction, the iTRAQ coupled with LC-MS/MS mass spectrometry and bioinformatics analysis were performed. We identified 288 differentially abundant proteins (DAPs) between yak and cattleyak epididymis; 151 were up-regulated while 137 were down-regulated in cattleyak relative to yak. Gene Ontology analysis identified that down-regulated DAPs in cattleyak were mostly enriched in the acetylation of protein component, along with negative and positive regulatory activities. iTRAQ proteomics data showed that the top up-regulated DAPs were mainly enriched in cell communication, cell adhesion, cytoskeleton organization, stress response, post-translational modifications and metabolic functions while the down-regulated DAPs were predominantly associated with sperm maturation, long-term sperm storage, sperm forward motility, sperm-oocyte fusion and regulatory functions. CONCLUSION: These results provide insight into the molecular mechanisms underlying male cattleyak sterility.

Analysis of long non-coding RNAs in epididymis of cattleyak associated with male infertility.

Cattleyak (CY), is a cross breed between cattle and yak (YK), which display equal adaptability to the harsh environment as YK and much higher performances than YK. However, the CY is female fertile and male sterile. Previous studies were conducted on testes tissues to investigate the mechanism of male infertility in CY. There is no systematic research on genes, especially lncRNAs between CY and YK epididymis. In this study, Illumina Hiseq was performed to profile the epididymis transcriptome (lncRNA and mRNA) of CY and YK. In total 18859 lncRNAs were identified, from which lincRNAs 12458, antisense lncRNAs 2345, intronic lncRNAs 3101, and sense lncRNAs 955 respectively. We have identified 345 DE lncRNAs and 3008 DE mRNAs between YK and CY epididymis. Thirteen DEGs were validated by quantitative real-time PCR. Combing with DEG, 14 couples of lncRNAs and their target genes were both DE, and 6 of them including CCDC39, KCNJ16, NECTIN2, MRPL20, PSMC4, and DEFB112 show their potential infertility-related terms such as cellular motility, sperm maturation, sperm storage, cellular junction, folate metabolism, and capacitation. On the other hand, several down-regulated genes such as DEFB124, DEFB126, DEFB125, DEFB127, DEFB129, CES5A, TKDP1, CST3, RNASE9 and CD52 in CY compared to YK were involved in the immune response and sperm maturation. Therefore, comprehensive analysis for lncRNAs and their target genes may enhance our understanding of the molecular mechanisms underlying the process of sperm maturation in CY and may provide important resources for further research.

ADIPOR1 regulates genes involved in milk fat metabolism in goat mammary epithelial cells.

BACKGROUND: Fat metabolism is a complex process regulated by a number of factors. Adiponectin receptor 1 (ADIPOR1) gene takes active part in lipid metabolism. Although, there have been some researches indicating that ADIPOR1 could influence the milk fat metabolism through targeting some factors, little is known about the effect of ADIPOR1 on goat milk fat metabolism. To investigate the regulatory role of ADIPOR1 on milk fat metabolism in GMECs, we analysed overexpression in the presence and absence of AdipoRon (50 µM) and examined knockdown using siRNA. Using RT-qPCR, we assessed ADIPOR1 mRNA expressions among different lactation stages in goat mammary gland and the expression of six genes that regulate milk fat metabolism in GMECs. RESULTS: ADIPOR1 mRNA expression level was higher during the various lactation stages, except dry-off period. Knockdown and overexpression results revealed a significant decrease and increase in mRNA expression of ADIPOR1 and genes considered: SREBF1, ACACA, FASN, SCD, ATGL, and HSL, respectively. Treatment of GMECs with AdipoRon 50 µM resulted in a significant (p < 0.05) increase in the mRNA expression of all measured genes, except SREBF1. CONCLUSION: Overall, ADIPOR1 plays a central role in regulating the transcription of several genes involved in milk fat metabolism.