Expression of <i>TXNRD1, HSPA4L</i> and <i>ATP1B1</i> Genes Associated with the Freezability of Boar Sperm
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 (<i>TXNRD1</i>), heat shock protein family A (HSP 70) member 4 like (<i>HSPA4L...
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MDPI AG
2022-08-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/16/9320 |
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| author | Anna Mańkowska Przemysław Gilun Łukasz Zasiadczyk Przemysław Sobiech Leyland Fraser |
| author_facet | Anna Mańkowska Przemysław Gilun Łukasz Zasiadczyk Przemysław Sobiech Leyland Fraser |
| author_sort | Anna Mańkowska |
| collection | DOAJ |
| description | 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 (<i>TXNRD1</i>), heat shock protein family A (HSP 70) member 4 like (<i>HSPA4L</i>) and sodium/potassium-transporting ATPase subunit beta-1 (<i>ATP1B1</i>) 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 <i>TXNRD1</i>, <i>HSPA4L</i> and <i>ATP1B1</i> 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 <i>TXNRD1</i> and <i>HSPA4L</i> in FT sperm compared with the fresh PF sperm. Furthermore, the relative mRNA expression level of <i>ATP1B1</i> 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 <i>ATP1B1</i> 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. |
| first_indexed | 2024-03-09T13:15:08Z |
| format | Article |
| id | doaj.art-0c6ecbb62dd044719abb3170accfe029 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T13:15:08Z |
| publishDate | 2022-08-01 |
| publisher | MDPI AG |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-0c6ecbb62dd044719abb3170accfe0292023-11-30T21:36:01ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-08-012316932010.3390/ijms23169320Expression of <i>TXNRD1, HSPA4L</i> and <i>ATP1B1</i> Genes Associated with the Freezability of Boar SpermAnna Mańkowska0Przemysław Gilun1Łukasz Zasiadczyk2Przemysław Sobiech3Leyland Fraser4Department of Animal Biochemistry and Biotechnology, Faculty of Animal Bioengineering, University of Warmia and Mazury in Olsztyn, 10-719 Olsztyn, PolandDepartment of Local Physiological Regulations, Institute of Animal Reproduction and Food Research of the Polish Academy of Sciences, Bydgoska 7, 10-243 Olsztyn, PolandDepartment of Animal Biochemistry and Biotechnology, Faculty of Animal Bioengineering, University of Warmia and Mazury in Olsztyn, 10-719 Olsztyn, PolandInternal Disease Unit, Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Warmia and Mazury in Olsztyn, 10-719 Olsztyn, PolandDepartment of Animal Biochemistry and Biotechnology, Faculty of Animal Bioengineering, University of Warmia and Mazury in Olsztyn, 10-719 Olsztyn, PolandCryopreservation 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 (<i>TXNRD1</i>), heat shock protein family A (HSP 70) member 4 like (<i>HSPA4L</i>) and sodium/potassium-transporting ATPase subunit beta-1 (<i>ATP1B1</i>) 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 <i>TXNRD1</i>, <i>HSPA4L</i> and <i>ATP1B1</i> 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 <i>TXNRD1</i> and <i>HSPA4L</i> in FT sperm compared with the fresh PF sperm. Furthermore, the relative mRNA expression level of <i>ATP1B1</i> 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 <i>ATP1B1</i> 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.https://www.mdpi.com/1422-0067/23/16/9320boarsemenfreezabilitystress-related genesproteins |
| spellingShingle | Anna Mańkowska Przemysław Gilun Łukasz Zasiadczyk Przemysław Sobiech Leyland Fraser Expression of <i>TXNRD1, HSPA4L</i> and <i>ATP1B1</i> Genes Associated with the Freezability of Boar Sperm International Journal of Molecular Sciences boar semen freezability stress-related genes proteins |
| title | Expression of <i>TXNRD1, HSPA4L</i> and <i>ATP1B1</i> Genes Associated with the Freezability of Boar Sperm |
| title_full | Expression of <i>TXNRD1, HSPA4L</i> and <i>ATP1B1</i> Genes Associated with the Freezability of Boar Sperm |
| title_fullStr | Expression of <i>TXNRD1, HSPA4L</i> and <i>ATP1B1</i> Genes Associated with the Freezability of Boar Sperm |
| title_full_unstemmed | Expression of <i>TXNRD1, HSPA4L</i> and <i>ATP1B1</i> Genes Associated with the Freezability of Boar Sperm |
| title_short | Expression of <i>TXNRD1, HSPA4L</i> and <i>ATP1B1</i> Genes Associated with the Freezability of Boar Sperm |
| title_sort | expression of i txnrd1 hspa4l i and i atp1b1 i genes associated with the freezability of boar sperm |
| topic | boar semen freezability stress-related genes proteins |
| url | https://www.mdpi.com/1422-0067/23/16/9320 |
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