6d1ae953c746d3b89c7a1318515b1b81 pmc_9311553.pdf 6f1c7b16593a05be1c08b26a9fe4b253801d361f pmc_9311553.pdf 2153e81ff088ab3c6784dae584cee413478ecb84e129b5322682af3f3318f8d2 pmc_9311553.pdf Title: Seminal Plasma Antioxidants Are Related to Sperm Cryotolerance in the Horse Subject: The objective of this study was to determine the relationship of enzymatic (superoxide dismutase, SOD; glutathione peroxidase, GPX; catalase, CAT; and paraoxonase type 1, PON1) and non-enzymatic antioxidants (measured in terms of: Trolox equivalent antioxidant capacity, TEAC; cupric-reducing antioxidant capacity, CUPRAC; and ferric-reducing ability of plasma, FRAP), as well as the oxidative stress index (OSI) in seminal plasma (SP) with the resilience of horse sperm to freeze-thawing. Twenty-one ejaculates (one per individual) were collected and split into two aliquots: the first was used to harvest the SP and assess the activity levels of antioxidants and the OSI, and the second one was cryopreserved. The following post-thaw sperm quality parameters were evaluated: sperm motility, plasma membrane and acrosome integrity, mitochondrial membrane potential, intracellular levels of reactive oxygen species (ROS), and plasma membrane lipid disorder. Based on post-thaw total motility (TM) and plasma membrane integrity (SYBR14+/PI-), ejaculates were hierarchically (p < 0.001) clustered into two groups of good (GFE) and poor (PFE) freezability. The SP activity levels of PON1, SOD, and TEAC were higher (p < 0.05) in GFE than in PFE, showing a positive relationship (p < 0.05) with some sperm motility parameters and with plasma membrane (PON1 and TEAC) and acrosome (SOD and TEAC) integrity. In contrast, OSI was higher (p < 0.05) in the SP of PFE than in that of GFE, and was negatively correlated (p < 0.05) to some sperm motility parameters and to plasma membrane and acrosome integrity, and positively (p < 0.05) to the percentage of viable sperm with high plasma membrane lipid disorder. In conclusion, enzymatic (PON1 and SOD) and non-enzymatic (TEAC) antioxidants of SP are related to horse sperm cryotolerance. In addition, our results suggest that PON1 could be one of the main antioxidant enzymes involved in the control of ROS in this species. Further investigation is needed to confirm the potential use of these SP-antioxidants and OSI to predict sperm cryotolerance in horses. Keywords: antioxidants; cryopreservation; horse; oxidative stress; reactive oxygen species; seminal plasma; sperm Author: Jaime Catalán, Iván Yánez-Ortiz, Asta Tvarijonaviciute, Luis Guillermo González-Aróstegui, Camila P. Rubio, Isabel Barranco, Marc Yeste and Jordi Miró Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Tue Jun 28 12:06:34 2022 CEST ModDate: Tue Jun 28 13:20:04 2022 CEST Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 20 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 3185354 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- JSIQSK+VnURWPalladioL Type 1 Custom yes yes no 10 0 DKPCYP+URWPalladioL-Bold Type 1 Custom yes yes no 15 0 YYSXZO+URWPalladioL-Roma Type 1 Custom yes yes no 20 0 REPYBZ+URWPalladioL-Ital Type 1 Custom yes yes no 24 0 PRBWYS+CMSY10 Type 1 Builtin yes yes no 28 0 ENXZUH+VnURWPalladioL-Bold Type 1 Custom yes yes no 32 0 KUDXZB+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 50 0 XKSBAY+EURM10 Type 1 Builtin yes yes no 75 0 BCDEEE+PalatinoLinotype-Roman TrueType WinAnsi 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BCDGEE+PalatinoLinotype-Italic TrueType WinAnsi yes yes no 308 0 BCDIEE+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 312 0 RCUAMB+VnURWPalladioL-Italic Type 1 Custom yes yes no 331 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2022-12-20 02:27:19 CET RepresentationInformation: pmc_9311553.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2022-12-14 02:18:02 CET Size: 3185354 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 691 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4923ab24 Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@44c8afef Children: Item: Title: Stallions and Samples Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7b69c6ba Item: Title: Seminal Plasma (SP) Collection Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@46daef40 Item: Title: Measurement of Enzymatic and Non-Enzymatic Antioxidant Levels in SP Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@12f41634 Item: Title: Measurement of Oxidative Stress Index (OSI) in SP Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@13c27452 Item: Title: Sperm Cryopreservation Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@262b2c86 Item: Title: Sperm Motility Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@371a67ec Item: Title: Sperm Functionality Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@5ed828d Children: Item: Title: Plasma Membrane Integrity (SYBR14/PI) Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@50d0686 Item: Title: Acrosome Integrity (PNA-FITC/PI) Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7a3d45bd Item: Title: Mitochondrial Membrane Potential (JC-1) Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1e7c7811 Item: Title: Intracellular Reactive Oxygen Species: Total ROS (H2DCFDA/PI) and O2- (HE/YO-PRO-1) Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77ec78b9 Item: Title: Plasma Membrane Lipid Disorder (M540/YO-PRO-1) Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1a3869f4 Item: Title: Statistical Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@a38d7a3 Children: Item: Title: Classification of Ejaculates Based on Their Cryotolerance Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77f99a05 Item: Title: Comparison of Antioxidant Variables Measured in SP between Good (GFE) and Poor Freezability Ejaculates (PFE) Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@63440df3 Item: Title: Correlations of Antioxidant Variables with Post-Thaw Semen Parameters Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@3aeaafa6 Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@76a3e297 Children: Item: Title: Classification of Horse Ejaculates Based on Their Cryotolerance Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4d3167f4 Item: Title: Activity Levels of Enzymatic Antioxidants in the SP of Horse Ejaculates Classified as GFE and PFE Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@ed9d034 Item: Title: Activity Levels of Non-Enzymatic Antioxidants in the SP of Horse Ejaculates Classified as GFE and PFE Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@6121c9d6 Item: Title: Oxidative Stress Index in the SP of Horse Ejaculates Classified as GFE and PFE Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@87f383f Item: Title: Correlations between SP Antioxidants (Enzymatic, Non-Enzymatic and OSI) and Post-Thaw Sperm Motility Parameters Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4eb7f003 Item: Title: Correlations between SP Antioxidants (Enzymatic, Non-Enzymatic and OSI) Levels and Post-Thaw Sperm Functionality Parameters Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@eafc191 Item: Title: Correlations between Seminal Plasma Antioxidants (Enzymatic, Non-Enzymatic and OSI) in Horse Ejaculates Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@612fc6eb Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1060b431 Item: Title: Conclusions Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@612679d6 Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@11758f2a Info: Title: Seminal Plasma Antioxidants Are Related to Sperm Cryotolerance in the Horse Author: Jaime Catalán, Iván Yánez-Ortiz, Asta Tvarijonaviciute, Luis Guillermo González-Aróstegui, Camila P. Rubio, Isabel Barranco, Marc Yeste and Jordi Miró Subject: The objective of this study was to determine the relationship of enzymatic (superoxide dismutase, SOD; glutathione peroxidase, GPX; catalase, CAT; and paraoxonase type 1, PON1) and non-enzymatic antioxidants (measured in terms of: Trolox equivalent antioxidant capacity, TEAC; cupric-reducing antioxidant capacity, CUPRAC; and ferric-reducing ability of plasma, FRAP), as well as the oxidative stress index (OSI) in seminal plasma (SP) with the resilience of horse sperm to freeze-thawing. Twenty-one ejaculates (one per individual) were collected and split into two aliquots: the first was used to harvest the SP and assess the activity levels of antioxidants and the OSI, and the second one was cryopreserved. The following post-thaw sperm quality parameters were evaluated: sperm motility, plasma membrane and acrosome integrity, mitochondrial membrane potential, intracellular levels of reactive oxygen species (ROS), and plasma membrane lipid disorder. Based on post-thaw total motility (TM) and plasma membrane integrity (SYBR14+/PI-), ejaculates were hierarchically (p < 0.001) clustered into two groups of good (GFE) and poor (PFE) freezability. The SP activity levels of PON1, SOD, and TEAC were higher (p < 0.05) in GFE than in PFE, showing a positive relationship (p < 0.05) with some sperm motility parameters and with plasma membrane (PON1 and TEAC) and acrosome (SOD and TEAC) integrity. In contrast, OSI was higher (p < 0.05) in the SP of PFE than in that of GFE, and was negatively correlated (p < 0.05) to some sperm motility parameters and to plasma membrane and acrosome integrity, and positively (p < 0.05) to the percentage of viable sperm with high plasma membrane lipid disorder. In conclusion, enzymatic (PON1 and SOD) and non-enzymatic (TEAC) antioxidants of SP are related to horse sperm cryotolerance. In addition, our results suggest that PON1 could be one of the main antioxidant enzymes involved in the control of ROS in this species. Further investigation is needed to confirm the potential use of these SP-antioxidants and OSI to predict sperm cryotolerance in horses. 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