Abstract
Due to lower farrowing rate and reduced litter size with frozen-thawed semen, over 90% of artificial insemination (AI) is conducted using liquid stored boar semen. Although substantial progress has been made towards optimizing the cryopreservation protocols for boar sperm, the influencing factors and underlying mechanisms related to cryoinjury and freeze tolerance of boar sperm remain largely unknown. In this study, we report the differential expression of mRNAs and miRNAs between fresh and frozen-thawed boar sperm using high-throughput RNA sequencing. Our results showed that 567 mRNAs and 135 miRNAs were differentially expressed (DE) in fresh and frozen-thawed boar sperm. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses revealed that the majority of DE mRNAs were enriched in environmental information processing such as cytokine-cytokine receptor interactions, PI3K-Akt signaling, cell adhesion, MAPK, and calcium signaling pathways. Moreover, the targets of DE miRNAs were enriched in significant GO terms such as cell process, protein binding, and response to stimuli. In conclusion, we speculate that DE mRNAs and miRNAs are heavily involved in boar sperm response to environment stimuli, apoptosis, and metabolic activities. The differences in expression also reflect the various structural and functional changes in sperm during cryopreservation.
Highlights
Artificial insemination (AI) has arguably been the most vital management tool employed for improving herd productivity in modern animal farming
More than 99% of AI is mainly conducted using liquid stored semen incubated at 15–20 ◦C for 0–5 days; while no more than 1% of AI procedures are performed with post-thawed boar semen due to the poorer farrowing rate and lower litter size [2,3]
Based on known miRNAs and newly predicted miRNAs and gene sequence information of the Tracdorirteiospnoanldlyin, g qsRpeTc-iPesC, RmiRisanduaseadnd toRNvAalhidybartied wtheere guesende toexpprreedsicstiotnhe lteavrgeelts gqenueasnotiffied by high-thdroifufegrehnptiualtlyseeqxpureenssceidngm.iRONuArsq(RTTab-PleCSR5).rTehsuenlt, sGsOhoanwdeKdEtGhGatatnhaelytsrisenofdaollfpdreifdfiecraetendtitaalrgeextspression of mRNoAf Ds EanmdiRmNiARsNiAn sfrienshfraensdh farnozdenfr‐tohzaewne-dthbaowaresdpberomarwsepreercmonwduacstecdon(TsaibstleesnSt 6wainthd tSh7e). dWifeferential expression patterns observed in RNA-seq data (Table S8)
Summary
Artificial insemination (AI) has arguably been the most vital management tool employed for improving herd productivity in modern animal farming. We have reported profiles of significantly dysregulated lncRNA and mRNA in fresh and frozen-thawed giant panda sperm These lncRNAs and mRNAs are mainly involved in regulating responses to cold stress and apoptosis via mechanisms such as the integral component of membrane, calcium transport, and PI3K-Akt, p53, and cAMP signaling pathways [41]. In order to achieve a better understanding of the transcriptomic events and transcriptional regulatory mechanisms, we used the integrated approaches of high-throughput transcriptome and small RNA sequencing to explore the differential expression profiles of mRNAs and miRNAs in fresh and post-thaw boar sperm. We speculate that our findings may serve as a helpful tool for further elucidation of the underlying molecular mechanism relevant to boar sperm cryopreservation
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