Determination of double-and single-stranded DNA breaks in bovine sperm is predictive of their fertilizing capacity  

作　　者：Jordi Ribas‑Maynou Ariadna Delgado‑Bermúdez Yentel Mateo‑Otero Estel Vinolas Carlos O.Hidalgo WSteven Ward Marc Yeste 

机构地区：[1]Biotechnology of Animal and Human Reproduction(TechnoSperm),Faculty of Sciences,Institute of Food and Agricultural Technology,University of Girona,C/Maria Aurèlia Campany,69,ES‑17003 Girona,Spain. [2]Unit of Cell Biology,Department of Biology,Faculty of Sciences,University of Girona,ES‑17003 Girona,Spain. [3]Institute for Biogenesis Research,Department of Anatomy,Biochemistry&Physiology,John A.Burns School of Medicine,University of Hawaii at Manoa,Honolulu,HI 96822,USA. [4]Department of Animal Selection and Reproduction,The Regional Agri-Food Research and Development Service of Asturias(SERIDA),ES‑33394 Gijón,Spain. [5]Catalan Institution for Research and Advanced Studies(ICREA),ES‑08010 Barcelona,Spain.

出　　处：《Journal of Animal Science and Biotechnology》2023年第1期134-151,共18页畜牧与生物技术杂志（英文版）

基　　金：the European Union’s Horizon 2020 Research and Innovation scheme under the Marie Sklodowska-Curie grant agreement No.801342(Tecniospring INDUSTRY,TECSPR-19-1-0003);the Ministry of Science and Innovation,Spain(AGL2017-88329-R and PID2020-113320RBI00);the Catalan Agency for Management of University and Research Grants,Regional Government of Catalonia,Spain(2017-SGR-1229);the Catalan Institution for Research and Advanced Studies(ICREA);La Maratóde TV3 Foundation(214/857-202039)。

摘　　要：Background:The analysis of chromatin integrity has become an important determinant of sperm quality.In frozenthawed bovine sperm,neither the sequence of post-thaw injury events nor the dynamics of different types of sperm DNA breaks are well understood.The aim of the present work was to describe such sperm degradation aftermath focusing on DNA damage dynamics,and to assess if this parameter can predict pregnancy rates in cattle.Results:A total of 75 cryopreserved ejaculates from 25 Holstein bulls were evaluated at two post-thawing periods(0-2 h and 2-4 h),analyzing global and double-stranded DNA damage through alkaline and neutral Comet assays,chromatin deprotamination and decondensation,sperm motility,viability,acrosomal status,and intracellular levels of total ROS,superoxides and calcium.Insemination of 59,605 females was conducted using sperm from the same bulls,thus obtaining the non-return to estrus rates after 90 d(NRR).Results showed an increased rate of double-stranded breaks in the first period(0-2 h:1.29±1.01%/h vs.2-4 h:0.13±1.37%/h;P<0.01),whereas the rate of sperm with moderate+high single-stranded breaks was higher in the second period(0-2 h:3.52±7.77%/h vs.2-4h:21.06±11.69%/h;P<0.0001).Regarding sperm physiology,viability decrease rate was different between the two periods(0-2 h:-4.49±1.79%/h vs.2-4 h:-2.50±3.39%/h;P=0.032),but the progressive motility decrease rate was constant throughout post-thawing incubation(0-2 h:-4.70±3.42%/h vs.2-4 h:-1.89±2.97%/h;P>0.05).Finally,whereas no correlations between bull fertility and any dynamic parameter were found,there were correlations between the NRR and the basal percentage of highly-damaged sperm assessed with the alkaline Comet(Rs=-0.563,P=0.003),between NRR and basal progressive motility(Rs=0.511,P=0.009),and between NRR and sperm with high ROS at 4 h post-thaw(Rs=0.564,P=0.003).Conclusion:The statistically significant correlations found between intracellular ROS,sperm viability,sperm motility,DNA damage and chromatin deprotamination suggested

关 键 词：Cattle CHROMATIN Comet test DNA damage FERTILITY SPERM Sperm quality 

分 类 号：S823[农业科学—畜牧学]