{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,17]],"date-time":"2026-10-17T21:10:34Z","timestamp":1792271434531,"version":"4.4.0"},"reference-count":81,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2022,10,5]],"date-time":"2022-10-05T00:00:00Z","timestamp":1664928000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001602","name":"Science Foundation Ireland","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001602","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002081","name":"Irish Research Council","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100002081","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Vet. Sci."],"abstract":"<jats:p>Bulls used in artificial insemination, with apparently normal semen quality, can vary significantly in their field fertility. This study aimed to characterize the transcriptome of spermatozoa from high (HF) and low (LF) fertility bulls at the mRNA and miRNA level in order to identify potential novel markers of fertility. Holstein-Friesian bulls were assigned to either the HF or LF group (<jats:italic>n<\/jats:italic> = 10 per group) based on an adjusted national fertility index from a minimum of 500 inseminations. Total RNA was extracted from a pool of frozen-thawed spermatozoa from three different ejaculates per bull, following which mRNA-seq and miRNA-seq were performed. Six mRNAs and 13 miRNAs were found differentially expressed (<jats:italic>P<\/jats:italic> &amp;lt; 0.05, FC &amp;gt; 1.5) between HF and LF bulls. Of particular interest, the gene pathways targeted by the 13 differentially expressed miRNAs were related to embryonic development and gene expression regulation. Previous studies reported that disruptions to protamine 1 mRNA (<jats:italic>PRM1<\/jats:italic>) had deleterious consequences for sperm chromatin structure and fertilizing ability. Notably, <jats:italic>PRM1<\/jats:italic> exhibited a higher expression in spermatozoa from LF than HF bulls. In contrast, Western Blot analysis revealed a decrease in PRM1 protein abundance for spermatozoa from LF bulls; this was not associated with increased protamine deficiency (measured by the degree of chromatin compaction) or DNA fragmentation, as assessed by flow cytometry analyses. However, protamine deficiency was positively and moderately correlated with the percentage of spermatozoa with DNA fragmentation, irrespective of fertility group. This study has identified potential biomarkers that could be used for improving semen quality assessments of bull fertility.<\/jats:p>","DOI":"10.3389\/fvets.2022.993561","type":"journal-article","created":{"date-parts":[[2022,10,5]],"date-time":"2022-10-05T06:09:39Z","timestamp":1664950179000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":23,"title":["Identification of differentially expressed mRNAs and miRNAs in spermatozoa of bulls of varying fertility"],"prefix":"10.3389","volume":"9","author":[{"given":"Eimear M.","family":"Donnellan","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jean-Philippe","family":"Perrier","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kate","family":"Keogh","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miriam","family":"\u0160tiavnick\u00e1","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Caitr\u00edona M.","family":"Collins","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elaine M.","family":"Dunleavy","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eli","family":"Sellem","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Naomi C.","family":"Bernecic","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patrick","family":"Lonergan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David A.","family":"Kenny","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sean","family":"Fair","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2022,10,5]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"433","DOI":"10.3168\/jds.2008-1646","article-title":"Invited review: Genomic selection in dairy cattle: progress and challenges","volume":"92","author":"Hayes","year":"2009","journal-title":"J Dairy Sci."},{"key":"B2","doi-asserted-by":"publisher","first-page":"2473","DOI":"10.3168\/jds.S0022-0302(00)75139-3","article-title":"Can relative spermatozoal galactosyltransferase activity be predictive of dairy bull fertility","volume":"83","author":"Larson","year":"2000","journal-title":"J Dairy Sci."},{"key":"B3","doi-asserted-by":"publisher","first-page":"1447","DOI":"10.1016\/j.theriogenology.2015.07.035","article-title":"Use of combinations of in vitro quality assessments to predict fertility of bovine semen","volume":"84","author":"Sellem","year":"2015","journal-title":"Theriogenology"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.3168\/jds.2021-20319","article-title":"Comprehensive functional analysis reveals that acrosome integrity and viability are key variables distinguishing artificial insemination bulls of varying fertility","author":"Bernecic","year":"2021","journal-title":"J Dairy Sci."},{"key":"B5","doi-asserted-by":"publisher","first-page":"24","DOI":"10.1016\/j.theriogenology.2020.07.005","article-title":"Differences in sperm functionality and intracellular metabolites in Norwegian Red bulls of contrasting fertility","volume":"157","author":"Narud","year":"2020","journal-title":"Theriogenology."},{"key":"B6","doi-asserted-by":"publisher","first-page":"42392","DOI":"10.1038\/srep42392","article-title":"Occurrence and functional significance of the transcriptome in bovine (Bos taurus) spermatozoa","volume":"7","author":"Selvaraju","year":"2017","journal-title":"Sci Rep."},{"key":"B7","doi-asserted-by":"publisher","first-page":"19","DOI":"10.1186\/s13072-020-00340-0","article-title":"comprehensive overview of bull sperm-borne small non-coding RNAs and their diversity across breeds","volume":"13","author":"Sellem","year":"2020","journal-title":"Epigenetics Chromatin."},{"key":"B8","doi-asserted-by":"publisher","first-page":"49","DOI":"10.1095\/biolreprod.112.103788","article-title":"Cryopreserved bovine spermatozoal transcript profile as revealed by high-throughput ribonucleic acid sequencing","volume":"88","author":"Card","year":"2013","journal-title":"Biol Reprod."},{"key":"B9","doi-asserted-by":"publisher","first-page":"570","DOI":"10.1016\/j.theriogenology.2011.03.009","article-title":"Association of CRISP2, CCT8, PEBP1 mRNA abundance in sperm and sire conception rate in Holstein bulls","volume":"76","author":"Arangasamy","year":"2011","journal-title":"Theriogenology."},{"key":"B10","doi-asserted-by":"publisher","first-page":"2007","DOI":"10.1016\/j.theriogenology.2012.07.016","article-title":"Association between mRNA abundance of functional sperm function proteins and fertility of Holstein bulls","volume":"78","author":"Kasimanickam","year":"2012","journal-title":"Theriogenology"},{"key":"B11","doi-asserted-by":"publisher","first-page":"256","DOI":"10.1016\/j.rvsc.2012.09.001","article-title":"Differential expression of protamine 1 and 2 genes in mature spermatozoa of normal and motility impaired semen producing crossbred Frieswal (HFxSahiwal) bulls","volume":"94","author":"Ganguly","year":"2013","journal-title":"Res Vet Sci."},{"key":"B12","doi-asserted-by":"publisher","first-page":"766","DOI":"10.1016\/j.theriogenology.2012.12.001","article-title":"Associations of adiponectin and fertility estimates in Holstein bulls","volume":"79","author":"Kasimanickam","year":"2013","journal-title":"Theriogenology."},{"key":"B13","doi-asserted-by":"publisher","first-page":"41","DOI":"10.1016\/j.theriogenology.2017.04.042","article-title":"Spermatozoal transcripts expression levels are predictive of semen quality and conception rate in bulls (Bos taurus)","volume":"98","author":"Parthipan","year":"2017","journal-title":"Theriogenology."},{"key":"B14","doi-asserted-by":"publisher","first-page":"162","DOI":"10.1016\/j.theriogenology.2016.11.011","article-title":"Associations of hypoosmotic swelling test, relative sperm volume shift, aquaporin7 mRNA abundance and bull fertility estimates","volume":"89","author":"Kasimanickam","year":"2017","journal-title":"Theriogenology."},{"key":"B15","doi-asserted-by":"publisher","first-page":"252","DOI":"10.1016\/j.theriogenology.2018.04.001","article-title":"Transcriptional abundance of type-1 endocannabinoid receptor (CB1) and fatty acid amide hydrolase (FAAH) in bull spermatozoa: Relationship with field fertility","volume":"114","author":"Kumar","year":"2018","journal-title":"Theriogenology."},{"key":"B16","doi-asserted-by":"publisher","first-page":"105","DOI":"10.1016\/j.anireprosci.2016.12.011","article-title":"Oligo-dT selected spermatozoal transcript profiles differ among higher and lower fertility dairy sires","volume":"177","author":"Card","year":"2017","journal-title":"Anim Reprod Sci."},{"key":"B17","doi-asserted-by":"publisher","first-page":"31794","DOI":"10.1038\/srep31794","article-title":"Characterisation of mouse epididymosomes reveals a complex profile of microRNAs and a potential mechanism for modification of the sperm epigenome","volume":"6","author":"Reilly","year":"2016","journal-title":"Sci Rep."},{"key":"B18","doi-asserted-by":"publisher","first-page":"635","DOI":"10.1242\/dev.131755","article-title":"Sperm-borne miRNAs and endo-siRNAs are important for fertilization and preimplantation embryonic development","volume":"143","author":"Yuan","year":"2016","journal-title":"Development."},{"key":"B19","doi-asserted-by":"publisher","first-page":"20347","DOI":"10.1038\/s41598-019-55868-3","article-title":"Characterization and functional roles of paternal RNAs in 2-4 cell bovine embryos","volume":"9","author":"Gross","year":"2019","journal-title":"Sci Rep."},{"key":"B20","doi-asserted-by":"publisher","first-page":"587","DOI":"10.1111\/rda.12531","article-title":"Expression of miRNAs in bull spermatozoa correlates with fertility rates","volume":"50","author":"Fagerlind","year":"2015","journal-title":"Reprod Domest Anim."},{"key":"B21","doi-asserted-by":"publisher","first-page":"e13412","DOI":"10.1111\/and.13412","article-title":"Sperm miR-15a and miR-29b are associated with bull fertility","volume":"52","author":"Menezes","year":"2020","journal-title":"Andrologia."},{"key":"B22","doi-asserted-by":"publisher","first-page":"14","DOI":"10.1186\/s12864-016-3394-7","article-title":"Small RNA sequencing of cryopreserved semen from single bull revealed altered miRNAs and piRNAs expression between High- and Low-motile sperm populations","volume":"18","author":"Capra","year":"2017","journal-title":"BMC Genomics."},{"key":"B23","doi-asserted-by":"publisher","first-page":"703101","DOI":"10.3389\/fvets.2021.703101","article-title":"Combined flow cytometric semen analysis and miRNA profiling as a tool to discriminate between high- and low-fertility bulls","volume":"8","author":"Turri","year":"2021","journal-title":"Front Vet Sci."},{"key":"B24","doi-asserted-by":"publisher","first-page":"165","DOI":"10.1186\/s12864-020-6570-8","article-title":"MicroRNA expression profile analysis in sperm reveals hsa-mir-191 as an auspicious omen of in vitro fertilization","volume":"21","author":"Xu","year":"2020","journal-title":"BMC Genomics."},{"key":"B25","doi-asserted-by":"publisher","first-page":"795","DOI":"10.1016\/j.theriogenology.2011.09.002","article-title":"Impact of genomic selection of AI dairy sires on their likely utilization and methods to estimate fertility: a paradigm shift","volume":"77","author":"Amann","year":"2012","journal-title":"Theriogenology."},{"key":"B26","doi-asserted-by":"publisher","first-page":"172","DOI":"10.1016\/j.theriogenology.2010.08.002","article-title":"Evaluation of bull fertility in dairy and beef cattle using cow field data","volume":"75","author":"Berry","year":"2011","journal-title":"Theriogenology."},{"key":"B27","doi-asserted-by":"publisher","first-page":"161","DOI":"10.1080\/01485010701216526","article-title":"The preparation of human spermatozoal RNA for clinical analysis","volume":"53","author":"Goodrich","year":"2007","journal-title":"Arch Androl."},{"key":"B28","doi-asserted-by":"publisher","first-page":"32","DOI":"10.1016\/j.ab.2015.03.022","article-title":"Spermatozoa input concentrations and RNA isolation methods on RNA yield and quality in bull (Bos taurus)","volume":"482","author":"Parthipan","year":"2015","journal-title":"Anal Biochem."},{"key":"B29","doi-asserted-by":"publisher","first-page":"374","DOI":"10.1111\/andr.12471","article-title":"High-quality human and rat spermatozoal RNA isolation for functional genomic studies","volume":"6","author":"Bianchi","year":"2018","journal-title":"Andrology."},{"key":"B30","doi-asserted-by":"publisher","first-page":"385","DOI":"10.1007\/978-1-62703-038-0_33","article-title":"Isolating mRNA and small noncoding RNAs from human sperm","volume":"927","author":"Goodrich","year":"2013","journal-title":"Methods Mol Biol."},{"key":"B31","doi-asserted-by":"publisher","first-page":"49","DOI":"10.1111\/and.12767","article-title":"Sperm RNA preparation for transcriptomic analysis: review of the techniques and personal experience","author":"El Fekih","year":"2017","journal-title":"Andrologia"},{"key":"B32","doi-asserted-by":"publisher","first-page":"772","DOI":"10.1016\/S0140-6736(02)09899-9","article-title":"Spermatozoal RNA profiles of normal fertile men","volume":"360","author":"Ostermeier","year":"2002","journal-title":"Lancet."},{"key":"B33","doi-asserted-by":"publisher","first-page":"134","DOI":"10.1186\/1471-2105-13-134","article-title":"Primer-BLAST: a tool to design target-specific primers for polymerase chain reaction","volume":"13","author":"Ye","year":"2012","journal-title":"BMC Bioinformatics."},{"key":"B34","doi-asserted-by":"publisher","first-page":"404","DOI":"10.1186\/s12864-018-4764-0","article-title":"A multi-scale analysis of bull sperm methylome revealed both species peculiarities and conserved tissue-specific features","volume":"19","author":"Perrier","year":"2018","journal-title":"BMC Genomics."},{"key":"B35","doi-asserted-by":"publisher","first-page":"2114","DOI":"10.1093\/bioinformatics\/btu170","article-title":"Trimmomatic: a flexible trimmer for Illumina sequence data","volume":"30","author":"Bolger","year":"2014","journal-title":"Bioinformatics."},{"key":"B36","doi-asserted-by":"publisher","first-page":"R36","DOI":"10.1186\/gb-2013-14-4-r36","article-title":"TopHat2: accurate alignment of transcriptomes in the presence of insertions, deletions and gene fusions","volume":"14","author":"Kim","year":"2013","journal-title":"Genome Biol."},{"key":"B37","doi-asserted-by":"publisher","first-page":"166","DOI":"10.1093\/bioinformatics\/btu638","article-title":"HTSeq\u2013a Python framework to work with high-throughput sequencing data","volume":"31","author":"Anders","year":"2015","journal-title":"Bioinformatics."},{"key":"B38","unstructured":"R: A language and environment for statistical computingViennaR Foundation for Statistical Computing2020"},{"key":"B39","doi-asserted-by":"publisher","first-page":"139","DOI":"10.1093\/bioinformatics\/btp616","article-title":"edgeR: a Bioconductor package for differential expression analysis of digital gene expression data","volume":"26","author":"Robinson","year":"2010","journal-title":"Bioinformatics."},{"key":"B40","unstructured":"FastQC: A Quality Control Tool for High Throughput Sequence Data\n            AndrewsS\n          2010"},{"key":"B41","unstructured":"Cutadapt Removes Adapter Sequences From High-Throughput Sequencing Reads\n            MartinM\n          2011"},{"key":"B42","doi-asserted-by":"publisher","first-page":"37","DOI":"10.1093\/nar\/gkr688","article-title":"miRDeep2 accurately identifies known and hundreds of novel microRNA genes in seven animal clades","volume":"40","author":"Friedlander","year":"2012","journal-title":"Nucleic Acids Res."},{"key":"B43","doi-asserted-by":"publisher","first-page":"D155","DOI":"10.1093\/nar\/gky1141","article-title":"miRBase: from microRNA sequences to function","volume":"47","author":"Kozomara","year":"2019","journal-title":"Nucleic Acids Res."},{"key":"B44","doi-asserted-by":"publisher","first-page":"R25","DOI":"10.1186\/gb-2009-10-3-r25","article-title":"Ultrafast and memory-efficient alignment of short DNA sequences to the human genome","volume":"10","author":"Langmead","year":"2009","journal-title":"Genome Biol."},{"key":"B45","doi-asserted-by":"publisher","first-page":"e05005","DOI":"10.7554\/eLife.05005","article-title":"Predicting effective microRNA target sites in mammalian mRNAs","volume":"4","author":"Agarwal","year":"2015","journal-title":"Elife."},{"key":"B46","doi-asserted-by":"publisher","first-page":"82","DOI":"10.1186\/1477-7827-10-82","article-title":"Dynamics of microRNAs in bull spermatozoa","volume":"10","author":"Govindaraju","year":"2012","journal-title":"Reprod Biol Endocrinol."},{"key":"B47","doi-asserted-by":"publisher","first-page":"201","DOI":"10.1006\/abio.1993.1104","article-title":"Rapid analysis of mammalian sperm nuclear proteins","volume":"209","author":"de Yebra","year":"1993","journal-title":"Anal Biochem."},{"key":"B48","doi-asserted-by":"publisher","first-page":"23","DOI":"10.1111\/j.1365-2605.1996.tb00429.x","article-title":"Chromomycin A3-staining as an indicator of protamine deficiency and fertilization","volume":"19","author":"Lolis","year":"1996","journal-title":"Int J Androl."},{"key":"B49","doi-asserted-by":"publisher","first-page":"370","DOI":"10.1111\/j.2047-2927.2014.00196.x","article-title":"Sperm protamine deficiency correlates with sperm DNA damage in Bos indicus bulls","volume":"2","author":"Fortes","year":"2014","journal-title":"Andrology."},{"key":"B50","doi-asserted-by":"publisher","first-page":"169","DOI":"10.1023\/A:1009844109023","article-title":"Sperm chromatin structure assay is useful for fertility assessment","volume":"22","author":"Evenson","year":"2000","journal-title":"Methods Cell Sci."},{"key":"B51","doi-asserted-by":"publisher","first-page":"27","DOI":"10.1093\/nar\/28.1.27","article-title":"kyoto encyclopedia of genes and genomes","volume":"28","author":"Kanehisa","year":"2000","journal-title":"Nucleic Acids Res."},{"key":"B52","doi-asserted-by":"publisher","first-page":"94","DOI":"10.1016\/j.theriogenology.2018.09.010","article-title":"Identification and differential expression of microRNAs in the testis of chicken with high and low sperm motility","volume":"122","author":"Liu","year":"2018","journal-title":"Theriogenology."},{"key":"B53","doi-asserted-by":"publisher","first-page":"539","DOI":"10.4103\/aja.aja_40_18","article-title":"Region-specific microRNA signatures in the human epididymis","volume":"20","author":"Browne","year":"2018","journal-title":"Asian J Androl."},{"key":"B54","doi-asserted-by":"publisher","first-page":"163","DOI":"10.1038\/nri2957","article-title":"MicroRNAs: the fine-tuners of Toll-like receptor signalling","volume":"11","author":"O'Neill","year":"2011","journal-title":"Nat Rev Immunol."},{"key":"B55","doi-asserted-by":"publisher","first-page":"853","DOI":"10.1093\/humrep\/dev031","article-title":"Serum miR-155 as a potential biomarker of male fertility","volume":"30","author":"Tsatsanis","year":"2015","journal-title":"Hum Reprod."},{"key":"B56","doi-asserted-by":"publisher","first-page":"54416","DOI":"10.18632\/oncotarget.17081","article-title":"Analysis of Bos taurus and Sus scrofa X and Y chromosome transcriptome highlights reproductive driver genes","volume":"8","author":"Khan","year":"2017","journal-title":"Oncotarget."},{"key":"B57","doi-asserted-by":"publisher","first-page":"77","DOI":"10.1111\/aji.12673","article-title":"MicroRNAs downregulated following immune activation of rat testis","author":"Parker","year":"2017","journal-title":"Am J Reprod Immunol"},{"key":"B58","doi-asserted-by":"publisher","first-page":"574","DOI":"10.1016\/j.febslet.2013.12.026","article-title":"MiR-98 is involved in rat embryo implantation by targeting Bcl-xl","volume":"588","author":"Xia","year":"2014","journal-title":"FEBS Lett."},{"key":"B59","doi-asserted-by":"publisher","first-page":"213","DOI":"10.1016\/j.fertnstert.2014.03.040","article-title":"New insights into the expression profile and function of micro-ribonucleic acid in human spermatozoa","volume":"102","author":"Salas-Huetos","year":"2014","journal-title":"Fertil Steril"},{"key":"B60","doi-asserted-by":"publisher","first-page":"1722","DOI":"10.1373\/clinchem.2011.169714","article-title":"Altered profile of seminal plasma microRNAs in the molecular diagnosis of male infertility","volume":"57","author":"Wang","year":"2011","journal-title":"Clin Chem."},{"key":"B61","doi-asserted-by":"publisher","first-page":"19130","DOI":"10.1038\/s41598-020-76152-9","article-title":"Plasma extracellular vesicle miRNAs as potential biomarkers of superstimulatory response in cattle","volume":"10","author":"Gad","year":"2020","journal-title":"Sci Rep."},{"key":"B62","doi-asserted-by":"publisher","first-page":"e106795","DOI":"10.1371\/journal.pone.0106795","article-title":"The expression pattern of microRNAs in granulosa cells of subordinate and dominant follicles during the early luteal phase of the bovine estrous cycle","volume":"9","author":"Salilew-Wondim","year":"2014","journal-title":"PLoS One."},{"key":"B63","doi-asserted-by":"publisher","first-page":"647717","DOI":"10.3389\/fcell.2021.647717","article-title":"Comparative Transcriptomic Analysis of Spermatozoa From High- and Low-Fertile Crossbred Bulls: Implications for Fertility Prediction","volume":"9","author":"Prakash","year":"2021","journal-title":"Front Cell Dev Biol."},{"key":"B64","doi-asserted-by":"publisher","first-page":"254","DOI":"10.1093\/humupd\/dmq035","article-title":"The different shades of mammalian pluripotent stem cells","volume":"17","author":"Kuijk","year":"2011","journal-title":"Hum Reprod Update."},{"key":"B65","doi-asserted-by":"publisher","first-page":"R173","DOI":"10.1530\/REP-19-0057","article-title":"The mTOR pathway in reproduction: from gonadal function to developmental coordination","volume":"159","author":"Correia","year":"2020","journal-title":"Reproduction."},{"key":"B66","doi-asserted-by":"publisher","first-page":"2121","DOI":"10.1007\/s10815-019-01555-1","article-title":"FoxO transcription factors 1 regulate mouse preimplantation embryo development","volume":"36","author":"Kuscu","year":"2019","journal-title":"J Assist Reprod Genet."},{"key":"B67","doi-asserted-by":"publisher","first-page":"34","DOI":"10.3390\/biology6030034","article-title":"MicroRNA signaling in embryo development","volume":"6","author":"Gross","year":"2017","journal-title":"Biology"},{"key":"B68","doi-asserted-by":"publisher","first-page":"502","DOI":"10.1111\/age.12941","article-title":"Integration of whole-genome DNA methylation data with RNA sequencing data to identify markers for bull fertility","volume":"51","author":"Gross","year":"2020","journal-title":"Anim Genet."},{"key":"B69","doi-asserted-by":"publisher","first-page":"106621","DOI":"10.1016\/j.anireprosci.2020.106621","article-title":"RNA-Seq analysis reveals functionally relevant coding and non-coding RNAs in crossbred bull spermatozoa","volume":"222","author":"Prakash","year":"2020","journal-title":"Anim Reprod Sci."},{"key":"B70","doi-asserted-by":"publisher","first-page":"92","DOI":"10.1095\/biolreprod.114.124255","article-title":"Sperm protamine-status correlates to the fertility of breeding bulls","volume":"92","author":"Dogan","year":"2015","journal-title":"Biol Reprod."},{"key":"B71","doi-asserted-by":"publisher","first-page":"151","DOI":"10.1002\/dvg.20598","article-title":"Genetic vasectomy-overexpression of Prm1-EGFP fusion protein in elongating spermatids causes dominant male sterility in mice","volume":"48","author":"Haueter","year":"2010","journal-title":"Genesis."},{"key":"B72","first-page":"S109","article-title":"Evaluation of protamines 1 and 2 transcript contents in spermatozoa from asthenozoospermic men","volume":"1","author":"Kempisty","year":"2007","journal-title":"Folia Histochem Cytobiol."},{"key":"B73","doi-asserted-by":"publisher","first-page":"57","DOI":"10.1016\/S1642-431X(12)60077-1","article-title":"Impact of protamine transcripts and their proteins on the quality and fertilization ability of sperm and the development of preimplantation embryos","volume":"12","author":"Depa-Martynow","year":"2012","journal-title":"Reprod Biol."},{"key":"B74","doi-asserted-by":"publisher","first-page":"535","DOI":"10.1093\/molehr\/gah064","article-title":"Analysis and significance of mRNA in human ejaculated sperm from normozoospermic donors: relationship to sperm motility and capacitation","volume":"10","author":"Lambard","year":"2004","journal-title":"Mol Hum Reprod."},{"key":"B75","doi-asserted-by":"publisher","first-page":"1408","DOI":"10.1016\/j.fertnstert.2006.04.024","article-title":"Sperm protamine 1\/protamine 2 ratios are related to in vitro fertilization pregnancy rates and predictive of fertilization ability","volume":"86","author":"Aoki","year":"2006","journal-title":"Fertil Steril."},{"key":"B76","doi-asserted-by":"publisher","first-page":"41","DOI":"10.1093\/molehr\/gah258","article-title":"novel mechanism of protamine expression deregulation highlighted by abnormal protamine transcript retention in infertile human males with sperm protamine deficiency","volume":"12","author":"Aoki","year":"2006","journal-title":"Mol Hum Reprod."},{"key":"B77","doi-asserted-by":"publisher","first-page":"60","DOI":"10.1023\/A:1009406231811","article-title":"Chromomycin A3 staining as a useful tool for evaluation of male fertility","volume":"17","author":"Iranpour","year":"2000","journal-title":"J Assist Reprod Genet."},{"key":"B78","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1017\/S0967199419000595","article-title":"Sperm DNA fragmentation: causes and identification","volume":"28","author":"Hamilton","year":"2020","journal-title":"Zygote."},{"key":"B79","doi-asserted-by":"publisher","first-page":"1039","DOI":"10.1002\/mrd.22871","article-title":"Reactive oxygen species as mediators of sperm capacitation and pathological damage","volume":"84","author":"Aitken","year":"2017","journal-title":"Mol Reprod Dev."},{"key":"B80","doi-asserted-by":"publisher","first-page":"1289","DOI":"10.1016\/j.fertnstert.2003.09.063","article-title":"Sperm chromatin structure assay (SCSA) parameters are related to fertilization, blastocyst development, and ongoing pregnancy in in vitro fertilization and intracytoplasmic sperm injection cycles","volume":"81","author":"Virro","year":"2004","journal-title":"Fertil Steril."},{"key":"B81","doi-asserted-by":"publisher","first-page":"2402","DOI":"10.1093\/humrep\/deu228","article-title":"Paternal influence of sperm DNA integrity on early embryonic development","volume":"29","author":"Simon","year":"2014","journal-title":"Hum Reprod."}],"container-title":["Frontiers in Veterinary Science"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fvets.2022.993561\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,10,5]],"date-time":"2022-10-05T06:09:44Z","timestamp":1664950184000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fvets.2022.993561\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,10,5]]},"references-count":81,"alternative-id":["10.3389\/fvets.2022.993561"],"URL":"https:\/\/doi.org\/10.3389\/fvets.2022.993561","relation":{},"ISSN":["2297-1769"],"issn-type":[{"value":"2297-1769","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,10,5]]},"article-number":"993561"}}