Identification of proteomic markers for ram spermatozoa motility using a tandem mass tag (TMT) approach.

Sperm motility is crucial for ram fertility; however, differences in the proteome of sperm with high- and low- motility in rams (Ovis aries) has yet to be achieved. Thus, the aim of this study was to identify and characterize ram spermatozoa proteins with different abundances between high- and low- motility, and identify of proteomic markers for ram spermatozoa motility using tandem mass tag (TMT) protein labeling and LC-MS/MS. In this study, the abundance of 150 proteins in high-motility (HM) ram sperm was significantly different compared with low-motility (LM) sperm. Proteins involved in sperm motility, mitochondrial activity, and spermatogenesis showed higher abundance in HM ram spermatozoa; proteins involved in protein processing and spliceosome were more abundant in LM ram spermatozoa. In conclusion, Phosphatidylethanolamine binding protein 4 is a potential proteomic marker for ram sperm motility; CCTs/HSPs are hallmarks of the LM spermatozoa in rams. Our findings highlight the functional differences between HM and LM ejaculated spermatozoa and has identified candidate proteins of interest linked to sperm motility in rams. SIGNIFICANCE: Inadequate sperm motility is one of the most important reasons for male subfertility or infertility. In this study, we found that the abundance of 150 proteins in high-motility ram sperm was significantly different compared with low-motility sperm. Proteomic biomarkers were discovered to reflect the motility variation in ram spermatozoa; these biomarkers may assist in illuminating the molecular mechanisms underlying sperm motility. This study expands the potential direction for sperm quality screening and animal breeding.

Identification and characterization of microRNAs in the ovaries of multiple and uniparous goats (Capra hircus) during follicular phase.

BACKGROUND: Superior kidding rate is an important economic trait in production of meat goat, and ovulation rate is the precondition of kidding rate. MicroRNAs (miRNAs) play critical roles in almost all ovarian biological processes, including folliculogenesis, follicle development, follicle atresia, luteal development and regression. To find out the different ovarian activity and follicle recruitment with miRNA-mediated posttranscriptional regulation, the small RNAs expressed pattern in the ovarian tissues of multiple and uniparous Anhui White goats during follicular phase was analyzed using Solexa sequencing data. RESULTS: 1008 miRNAs co-expressed, 309 and 433 miRNAs specifically expressed in the ovaries of multiple and uniparous goats during follicular phase were identified. The 10 most highly expressed miRNAs in the multiple library were also the highest expressed in the uniparous library, and there were no significantly different between each other. The highest specific expressed miRNA in the multiple library was miR-29c, and the one in the uniparous library was miR-6406. 35 novel miRNAs were predicted in total. GO annotation and KEGG Pathway analyses were implemented on target genes of all miRNA in two libraries. RT-PCR was applied to detect the expression level of 5 randomly selected miRNAs in multiple and uniparous hircine ovaries, and the results were consistent with the Solexa sequencing data. CONCLUSIONS: In the present study, the different expression of miRNAs in the ovaries of multiple and uniparous goats during follicular phase were characterized and investigated using deep sequencing technology. The result will help to further understand the role of miRNAs in kidding rate regulation and also may help to identify miRNAs which could be potentially used to increase hircine ovulation rate and kidding rate in the future.