Expression of TXNRD1, HSPA4L and ATP1B1 Genes Associated with the Freezability of Boar Sperm

Abstract

Cryopreservation is associated with increased oxidative stress, which is responsible for sperm damage. We analyzed the effect of cryopreservation on mRNA and protein expression of thioredoxin reductase 1 (TXNRD1), heat shock protein family A (HSP 70) member 4 like (HSPA4L) and sodium/potassium-transporting ATPase subunit beta-1 (ATP1B1) genes in boar sperm with different freezability. Boars were classified as having good and poor semen freezability (GSF and PSF, respectively), according to the assessment of post-thaw sperm motility. Total RNA was isolated from fresh pre-freeze (PF) and frozen-thawed (FT) sperm from five boars of the GSF and PSF groups, respectively. Quantification of TXNRD1, HSPA4L and ATP1B1 gene expression was performed by RT-qPCR analysis. Proteins extracted from sperm were subjected to Western blotting and SDS-PAGE analyses. Poor freezability ejaculates were characterized by significantly higher relative mRNA expression levels of TXNRD1 and HSPA4L in FT sperm compared with the fresh PF sperm. Furthermore, the relative mRNA expression level of ATP1B1 was significantly higher in the fresh PF sperm of the GSF group. Western blotting analysis revealed significantly higher relative expression of TXNRD1 protein in the fresh PF sperm of the GSF group, while HSPA4L protein expression was markedly increased in FT sperm of the PSF group. Electrophoretic and densitometric analyses revealed a higher number of proteins in the fresh PF and FT sperm of the PSF and GSF groups, respectively. The results of this study indicate that ATP1B1 mRNA expression in the fresh PF sperm is a promising cryotolerance marker, while the variations of TXNRD1 and HSPA4L protein expression in the fresh PF or FT sperm provide useful information that may help to elucidate their biological significance in cryo-damage.

Extracellular vesicle-coupled miRNA profiles of chicken seminal plasma and their potential interaction with recipient cells

“…The high abundant SPEV-coupled miRNAs also target 107 functional mRNAs in the sperm. The heat shock protein family A member 4 like and eukaryotic translation elongation factor 1 beta 2 targeted by gga-miR-10b-5p and gga-miR-10a-5p, respectively, were highly expressed in hypospermate individuals and were associated with embryonic development ( Selvaraju et al, 2021 ; Mańkowska et al, 2022 ). The DEAD-box helicase 4 is a germline-specific gene and Transglutaminase 4, a prostate-specific gene used as a control gene, are also regulated by gga-miR-10b-5p and gga-miR-10a-5p ( Kim et al, 2015 ; Yu et al, 2016a ).…”

A Comprehensive Genomic Analysis of Chinese Indigenous Ningxiang Pigs: Genomic Breed Compositions, Runs of Homozygosity, and Beyond

“…The Ningxiang pigs, classified as an obese-type breed, are renowned for their abundant fat content and excellent meat quality. In this study, we identified genes ZNF280D [41], PLIN1 [42], PAPPA2 [43], SLC5A4 [44], and CABIN1 [45], which were found to be associated with BFT in pigs, and CCPG1 [46], ATP1B1 [47], and CABS1 [16], which demonstrated associations with reproductive performance in boars. Additionally, we identified some genes that simultaneously influence multiple traits.…”

Molecular Markers: A New Paradigm in the Prediction of Sperm Freezability

“…However, TXNRD1 should work as a scavenger of ROS by controlling the redox state of oxidized proteins; however, even when the expression was higher in the cryopreserved group, the potential cryo-induced oxidative damage may have decreased the quality of frozen boar spermatozoa after thawing. The ATP1B1 gene may be used as a cryotolerance marker of spermatozoa to cold shock because a significant loss of mRNA expression as well as ATP1B1 protein in fresh prefreeze samples predicts reduced freezability [ 62 ].…”