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Sperm function, mitochondrial activity and in vivo fertility are associated to their mitochondrial DNA content in pigs

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Journal of Animal Science and Biotechnology 2024年第4期15卷 1550-1558页

作者： Marc Llavanera Yentel Mateo‑Otero Estel Vinolas‑Vergés sergi bonet Marc Yeste Unit of Cell Biology Department of BiologyFaculty of SciencesUniversity of GironaGirona ES‑17003Spain Biotechnology of Animal and Human Reproduction(TechnoSperm) Institute of Food and Agricultural TechnologyUniversity of GironaGirona ES‑17003Spain Catalan Institution for Research and Advanced Studies(ICREA) Barcelona ES‑08010Spain

Background Despite their low abundance in sperm, mitochondria have diverse functions in this cell type, includ-ing energy production, signalling and calcium regulation. In humans, sperm mitochondrial DNA content(mtDNAc) has been reported to be negatively linked to sperm function and fertility. Yet, the association between mtDNAc and sperm function in livestock remains unexplored. For this reason, this study aimed to shed some light on the link between mtDNAc and sperm function and fertilising potential in pigs. A qPCR method for mtDNAc quantification was optimised for pig sperm, and the association of this parameter with sperm motility, kinematics, mitochondrial activity, and fertility was subsequently *** First, the q PCR method was found to be sensitive and efficient for mtDNAc quantification in pig sperm. By using this technique, mtDNAc was observed to be associated to sperm motility, mitochondrial activity and in vivo, but not in vitro, fertility outcomes. Specifically, sperm with low mtDNAc were seen to exhibit greater motility but decreased mitochondrial activity and intracellular reactive oxygen species. Interestingly, samples with lower mtD-NAc showed higher conception and farrowing rates, but similar in vitro fertilisation rates and embryo development, when compared to those with greater *** These findings enrich our comprehension of the association of mtDNAc with sperm biology, and lay the foundation for future research into employing this parameter as a molecular predictor for sperm function and fer-tility in livestock.

关键词： Fertility Mitochondria Motility mtDNA copy number qPCR Sperm

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Chromatin condensation but not DNA integrity of pig sperm is greater in the sperm-rich fraction

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Journal of Animal Science and Biotechnology 2024年第1期15卷 171-181页

作者： Estel Viñolas-Vergés Jordi Ribas-Maynou Isabel Barranco Camila Peres Rubio sergi bonet Jordi Roca Marc Yeste Unit of Cell Biology Department of BiologyFaculty of SciencesUniversity of GironaGironaSpain Biotechnology of Animal and Human Reproduction(TechnoSperm) Institute of Food and Agricultural TechnologyUniversity of GironaGironaSpain Department of Medicine and Animal Surgery Faculty of Veterinary ScienceUniversity of MurciaMurciaSpain Catalan Institution for Research and Advanced Studies(ICREA) S08010 BarcelonaSpain

Background Protamination and condensation of sperm chromatin as well as DNA integrity play an essential role during fertilization and embryo *** some mammals,like pigs,ejaculates are emitted in three separate fractions:pre-sperm,sperm-rich(SRF)and post sperm-rich(PSRF).These fractions are known to vary in volume,sperm concentration and quality,as well as in the origin and composition of seminal plasma(SP),with differences being also observed within the SRF ***,whether disparities in the DNA integrity and chromatin condensation and pro-tamination of their sperm exist has not been *** This study determined chromatin protamination(Chromomycin A3 test,CMA_(3)),condensation(Dibromobi-mane test,DBB),and DNA integrity(Comet assay)in the pig sperm contained in the first 10 m L of the SRF(SRF-P1),the remaining portion of the sperm-rich fraction(SRF-P2),and the post sperm-rich fraction(PSRF).While chromatin protamination was found to be similar between the different ejaculate fractions(P>0.05),chromatin condensation was seen to be greater in SRF-P1 and SRF-P2 than in the PSRF(P=0.018 and P=0.004,respectively).Regarding DNA integrity,no differences between fractions were observed(P>0.05).As the SRF-P1 has the highest sperm concentra-tion and ejaculate fractions are known to differ in antioxidant composition,the oxidative stress index(OSi)in SP,calcu-lated as total oxidant activity divided by total antioxidant capacity,was tested and confirmed to be higher in the SRF-P1 than in SRF-P2 and PSRF(0.42±0.06 vs.0.23±0.09 and 0.08±0.00,respectively;P<0.01);this index,in addition,was observed to be correlated to the sperm concentration of each fraction(Rs=0.973;P<0.001).Conclusion While sperm DNA integrity was not found to differ between ejaculate fractions,SRF-P1 and SRF-P2 were observed to exhibit greater chromatin condensation than the *** could be related to the OSi of each fraction.

关键词： Chromatin Condensation DNA integrity Ejaculate fractions Pig Protamination Sperm

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GSTM3, but not IZUMO1, is a cryotolerance marker of boar sperm

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Journal of Animal Science and Biotechnology 2019年第4期10卷 931-941页

作者： Marc Llavanera Ariadna Delgado-Bermúdez Beatriz Fernandez-Fuertes Sandra Recuero Yentel Mateo sergi bonet Isabel Barranco Marc Yeste Biotechnology of Animal and Human Reproduction (Techno Sperm) Unit of Cell Biology Department of Biology Institute of Food and Agricultural Technology University of Girona

Background: Cryopreservation is currently the most efficient method for long-term preservation of mammalian gametes and is extensively used in swine artificial insemination(AI) centres. However, it is well-known that cryopreservation procedures induce changes in the water phase in both intra and extracellular compartments,which alter the content and localisation of several proteins and ends up curtailing the structural integrity of functional sperm(i.e., cryoinjuries). Alterations and deficiencies of sperm-oocyte binding proteins during gamete recognition are one of the causes of reproductive failure both in vitro and in vivo. In this sense, characterisation of cryopreservation effects upon oocyte-binding proteins of sperm, such as IZUMO1 and GSTM3, is essential when assessing the impact of this technique in swine ***: Cryopreservation was found to induce changes in the localisation of IZUMO1 and GSTM3 in boar ***, the relative content of both proteins was not altered after thawing. Furthermore, whereas IZUMO1 content was found not to be related to the cryotolerance of boar sperm, GSTM3 content was observed to be higher in poor(PFE) than in good(GFE) freezability ejaculates in both pre-frozen(1.00 INT·mm^2± 0.14 INT·mm^2 vs.0.72 INT·mm^2± 0.15 INT·mm^2;P < 0.05) and post-thawed(0.96 INT·mm^2± 0.20 INT·mm^2 vs. 70 INT·mm^2± 0.19 INT·mm^2;P < 0.05) samples. Moreover, GSTM3 levels were found to be higher in those spermatozoa that exhibited low mitochondrial activity, high reactive oxygen species(ROS) production, and high membrane lipid disorder postthaw(P < 0.05).Conclusions: The difference in GSTM3 content between GFE and PFE, together with this protein having been found to be related to poor sperm quality post-thaw, suggests that it could be used as a cryotolerance marker of boar spermatozoa. Furthermore, both IZUMO1 and GSTM3 relocate during cryopreservation, which could contribute to the reduced fertilising capacity of frozen-thawed boar

关键词： Boar Cryopreservation GSTM3 IZUMO1 ROS Sperm

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Aquaglyceroporins but not orthodox aquaporins are involved in the cryotolerance of pig spermatozoa

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Journal of Animal Science and Biotechnology 2020年第1期11卷 20-31页

作者： Ariadna Delgado-Bermúdez Marc Llavanera Leira Fernández-Bastit Sandra Recuero Yentel Mateo-Otero sergi bonet Isabel Barranco Beatriz Fernández-Fuertes Marc Yeste Biotechnology of Animal and Human Reproduction(TechnoSperm) Unit of Cell BiologyDepartment of BiologyFaculty of SciencesInstitute of Food and Agricultural TechnologyUniversity of GironaC/Maria Aurèlia Campany69Campus MontiliviE-17003 GironaSpain

Background:Aquaporins(AQPs)are a family of transmembrane water channels that includes orthodox AQPs,aquaglyceroporins(GLPs)and super ***3,AQP7,AQP9 and AQP11 have been identified in boar sperm,and they are crucial for sperm maturation and *** exchange is an important event in cryopreservation,which is the most efficient method for long-term storage of ***,the freezethaw process leads to sperm damage and a loss of fertilizing *** that the quality of frozenthawed sperm partially depends on the regulation of osmolality variations during this process,AQPs might play a crucial role in boar semen *** this context,the aim of this study was to unravel the functional relevance of the different groups of AQPs for boar sperm cryotolerance through three different ***:Inhibition of different groups of AQPs was found to have different effects on boar sperm *** the use of 1,3-propanediol(PDO),an inhibitor of orthodox AQPs and GLPs,decreased total motility(P<0.05),it increased post-thaw sperm viability,lowered membrane lipid disorder and increased mitochondrial membrane potential(MMP)(P<0.05).When acetazolamide(AC)was used as an inhibitor of orthodox AQPs,the effects on post-thaw sperm quality were restricted to a mild increase in MMP in the presence of the intermediate concentration at 30 min post-thaw and an increase in superoxide levels(P<0.05).Finally,the addition of phloretin(PHL),a GLP inhibitor,had detrimental effects on post-thaw total and progressive sperm motilities,viability and lipid membrane disorder(P<0.05).Conclusions:The effects of the different inhibitors suggest that GLPs rather than orthodox AQPs are relevant for boar sperm ***,the positive effect of PDO on sperm quality suggests a cryoprotective role for this molecule.

关键词： Acetazolamide Aquaporins Boar Phloretin Propanediol Sperm

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Sperm chromatin condensation as an in vivo fertility biomarker in bulls:a flow cytometry approach

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Journal of Animal Science and Biotechnology 2022年第2期13卷 418-429页

作者： Marc Llavanera Jordi Ribas-Maynou Ariadna Delgado-Bermúdez Sandra Recuero Rodrigo Muiño Carlos OHidalgo Carolina Tamargo sergi bonet Yentel Mateo-Otero Marc Yeste Biotechnology of Animal and Human Reproduction(TechnoSperm) Faculty of SciencesUniversity of GironaC/Maria Aurèlia Campany69ES-17003 GironaSpain Department of Biology Unit of Cell BiologyFaculty of SciencesUniversity of GironaES-17003 GironaSpain Department of Animal Pathology Faculty of Veterinary MedicineUniversity of Santiago de CompostelaES-15705 LugoSpain Department of Animal Selection and Reproduction The Regional Agri-Food Research and Development Service of Asturias(SERIDA)E-33394 GijónSpain

Background:Genetic selection in cattle has been directed to increase milk ***,coupled to the fact that the vast majority of bovine artificial inseminations(AI)are performed using cryopreserved sperm,have led to a reduction of fertility rates over the ***,seeking sensitive and specific sperm biomarkers able to predict fertility rates is of vital importance to improve cattle reproductive *** humans,sperm chromatin condensation evaluated through chromomycin A3(CMA3)has recently been purported to be a powerful biomarker for sperm functional status and male *** objectives of the present study were:a)to set up a flow cytometry method for simultaneously evaluating chromatin condensation and sperm viability,and b)to test whether this parameter could be used as a predictor of in vivo fertility in *** study included pools of three independent cryopreserved ejaculates per bull from 25 Holstein *** outcomes of each sire were determined by non-return rates,which were used to classify bulls into two groups(highly fertile and subfertile).Results:Chromatin condensation status of bovine sperm was evaluated through the combination of CMA3 and Yo-Pro-1 staining and flow *** quality parameters(morphology,viability,total and progressive motility)were also *** correlation coefficients and ROC curves were calculated to assess their capacity to predict in vivo *** morphology,viability and total motility presented an area under the ROC curve(AUC)of 0.54,0.64 and 0.68,respectively(P>0.05),and thus were not able to discriminate between fertile and subfertile ***,while the percentage of progressively motile sperm showed a significant predictive value,with an AUC of 0.73(P=0.05),CMA3/Yo-Pro-1 staining even depicted superior results for the prediction of in vivo fertility in ***,the percentage of viable sperm with poor chromatin condensation showed better accura

关键词： Bull Chromatin Chromomycin A3 Condensation Fertility Flow cytometry Sperm

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Sperm function, mitochondrial activity and in vivo fertility are associated to their mitochondrial DNA content in pigs.

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畜牧与生物技术杂志(英文版) 2024年第4期 1550页

作者： Marc Llavanera Yentel Mateo-Otero Estel Vi?olas-Vergés sergi bonet Marc Yeste

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Chromatin condensation but not DNA integrity of pig sperm is greater in the sperm‑rich fraction

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畜牧与生物技术杂志(英文版) 2024年第1期

作者： Estel Viñolas-Vergés Jordi Ribas-Maynou Isabel Barranco Camila Peres Rubio sergi bonet Jordi Roca Marc Yeste

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GSTM3, but not IZUMO1, is a cryotolerance marker of boar sperm.

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畜牧与生物技术杂志(英文版) 2019年第4期

作者： Marc Llavanera Ariadna Delgado-Bermúdez Beatriz Fernandez-Fuertes Sandra Recuero Yentel Mateo sergi bonet Isabel Barranco and Marc Yeste Biotechnology of Animal and Human Reproduction (Techno Sperm)，Unit of Cell Biology，Department of Biology，Institute of Food and Agricultural Technology，University of Girona

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Sperm chromatin condensation as an in vivo fertility biomarker in bulls: a flow cytometry approach

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畜牧与生物技术杂志(英文版) 2022年第2期

作者： Marc Llavanera Jordi Ribas-Maynou Ariadna Delgado-Bermúdez Sandra Recuero Rodrigo Mui?o Carlos O.Hidalgo Carolina Tamargo sergi bonet Yentel Mateo-Otero Marc Yeste

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