Transcriptome analysis of boar spermatozoa with different freezability using RNA-Seq.

Semen freezability is associated with genetic markers, and there is a diverse set of sperm transcripts that have been attributed to various cellular functions. RNA-Seq was performed to compare the transcript profiles of spermatozoa from boars with different semen freezability. We examined ejaculates from the Polish large white (PLW) boars that were classified as having good and poor semen freezability (GSF and PSF, respectively; n = 3 boars per group) by assessing post-thaw motility characteristics, mitochondrial membrane potential, plasma membrane and acrosome integrity. Total RNA was isolated from fresh spermatozoa from boars of the GSF and PSF groups and subjected to RNA-Seq (Illumina NextSeq 500 platform). Transcript abundance was assessed with the DESeq2, DESeq, and EdgeR Bioconductor R packages, and varying numbers of differentially expressed gene (DEG) transcripts were detected in the spermatozoa of each boar. Using RNA-Seq, we identified several genes associated with inflammation and apoptosis (FOS, NFATC3, ITGAL, EAF2 and ZDHHC14), spermatogenesis (FGF-14 and BAMBI), autophagy (RAB33B), protein phosphorylation (PTPRU and PTPN2) and energy metabolism (ND6 and ACADM) that were predominantly up-regulated in poor freezability ejaculates. Quantitative reverse transcription polymerase chain reaction (RT-qPCR) validated the transcript expression levels detected by RNA-Seq and thus confirmed the reliability of this technique. Subsequent validation with western blotting showed that the expression of three proteins was in accordance with the transcript abundance. Overall, we demonstrated that the up-regulation of the DEG transcripts in spermatozoa was associated with poor semen freezability. We suggest that spermatozoa transcriptome profiling provides a foundation to further elucidate the relevance of sperm-related transcripts on cryo-survival. The sperm-related transcripts, namely FOS, NFATC3, EAF2, BAMBI, PTPRU, PTPN2, ND6 and ACADM, are potential markers for predicting the freezability of boar semen.

Sazonalidade na congelabilidade de sêmen bovino / Seasonality in the freezability of bovine semen / Estacionalidad en congelamiento de semen bovino

O objetivo desta revisão foi apresentar e discutir os efeitos da sazonalidade nas características seminais de touros Bos taurus e Bos indicus antes e após o congelação, a fim de possibilitar maior rentabilidade econômica. Apesar da reprodução da espécie bovina não se caracterizar pela sazonalidade, estudos mostram a interferência das estações do ano na qualidade do sêmen de touros. O aumento da temperatura ambiente pode levar a elevação da temperatura testicular, das taxas metabólicas e das exigências de oxigênio, ocasionando alterações na espermatogênese e, por consequência, a um impacto negativo na reprodução. Touros Bos indicus obtêm melhor desempenho na estação mais quente do ano apresentando uma qualidade de sêmen satisfatória em relação a touros Bos taurus. Isso se deve à maior resistência dos zebuínos às altas temperaturas e a peroxidação lipídica causada pelo calor resultando em menor produção de espécies reativas ao oxigênio e menor aumento nos defeitos nas células espermáticas. Desse modo, touros de origem europeia não apresentam um bom desempenho durante alguns períodos mais quentes do ano o que leva a questionar se a relação custo-benefício de aproveitamento do sêmen para criopreservação justifica o processamento do mesmo pelas Centrais de Coleta e Processamento de Sêmen nesta época. Esse fato tem levado algumas empresas a suspender atividades em determinados meses do ano.(AU)

The objective of this review was to present and discuss the effects of seasonality in the semen characteristics of Bos taurus and Bos indicus bulls before and after freezing in order to enable greater economic profitability. Even though the bovine reproduction is not characterized by seasonality, studies have shown the interference of the season on semen quality in bulls. The increase of temperature may lead to increase in testicular temperature, metabolic rates and oxygen requirements. Both thermal and oxidative stress will cause changes in the normal development of spermatogenesis, changing volume, sperm concentration, progressive motility, vigor and morphology of sperm cells, resulting in an economically negative impact on reproduction. Bos indicus bulls presented better performance in the hottest season of the year, showing a satisfactory quality of semen when compared to Bos taurus bulls. This is due to the greater resistance to high temperatures of zebu bulls and lipid peroxidation caused by the heat resulting in lower production of species reactive to oxygen and smaller increases in defects in sperm cells. Thus, European bulls, such as Bos Taurus, do not present a good performance in some of the hottest periods of the year, which leads to questioning if the cost-effective use of semen cryopreservation justifies its processing by the Semen Collection and Processing Centers during these periods. This fact has led some national centers for semen suspend activities in certain months of the year.(AU)

El objetivo de esta revisión ha sido presentar y discutir los efectos de la estacionalidad en las características del semen de toros Bos taurus y Bos indicus pre y post congelación, con el fin de proporcionar mayor rentabilidad económica. A pesar de la reproducción vacuna no caracterizarse por la estacionalidad, estudios muestran la interferencia de las estaciones del año en la calidad del semen de toros El aumento de la temperatura ambiente puede conducir a elevada temperatura testicular, de las tasas metabólicas y de las exigencias de oxígeno, ocasionando alteraciones en la espermatogénesis y, por lo tanto, a un impacto negativo en la reproducción. Toros Bos indicus presentaron mejor rendimiento en la temporada más calurosa del año, presentando calidad de semen satisfactoria en comparación con toros Bos taurus. Esto es debido a mayor resistencia del ganado cebú a las altas temperaturas y la peroxidación de lípidos causada por el calor, que resulta en menor producción de especies reactivas al oxígeno y menos defectos en las células espermáticas. Así, toros de origen europea no presentan buen desempeño durante algunos períodos más calurosos del año, lo que lleva a cuestionar si la relación costo beneficio de aprovechamiento del semen para criopreservación justifica la tramitación del mismo por las Centrales de Recogida y Procesamiento de Semen en este periodo. Este hecho ha llevado a algunas empresas a suspender las actividades en ciertos meses del año.(AU)