FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / Sci Rep / Proteomic analysis of sperm from fertile stallions and subfe...
Scientific reports2024; 14(1); 12446; doi: 10.1038/s41598-024-63410-3

Proteomic analysis of sperm from fertile stallions and subfertile stallions due to impaired acrosomal exocytosis.

Abstract: Thoroughbred stallions that carry a double-homozygous genotype A/A-A/A for SNPs rs397316122 and rs69101140 in exon 5 of the FKBP6 gene (chr13; EquCab3.0) are uniquely subfertile due to impaired acrosomal exocytosis (IAE). In this study, the sperm proteome in frozen/thawed semen from subfertile Thoroughbred stallions was studied and compared to that of frozen/thawed sperm from fertile Thoroughbred stallions. A total of 2,220 proteins was identified, of which 140 proteins were found to be differentially abundant in sperm from the subfertile stallions compared to that of fertile stallions (83 less and 57 more abundant). Proteins of differential abundance in sperm from the subfertile stallions were mainly overrepresented in the "metabolism" and the "metabolism of lipids" pathways. One of these proteins, arylsulfatase F (ARSF), was studied by immunofluorescence. A lower proportion of sperm displaying ARSF signal at the acrosome region was observed in sperm from subfertile Thoroughbred stallions. In addition, heterologous zona pellucida binding assays revealed that sperm from subfertile Thoroughbred stallions bound at a lower proportion to zonae pellucidae than sperm from fertile Thoroughbred stallions. In conclusion, a group of differential abundance proteins, including some of acrosome origin, were identified in sperm from subfertile stallions with acrosome dysfunction.
© 2024. The Author(s).
Get Full Text
Publication Date: 2024-05-30 PubMed ID: 38816557PubMed Central: PMC11139894DOI: 10.1038/s41598-024-63410-3Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This study focused on comparing the protein composition in the semen of fertile and subfertile Thoroughbred stallions, with a particular interest in those suffering from impaired acrosomal exocytosis (IAE), a condition associated with two specific genotypes. Results highlighted significant differences in certain proteins and pathways suggesting a potential connection with the fertility issues.

Comparison of Sperm Proteome in Fertile and Subfertile Stallions

  • The primary goal of the research was to investigate the differences in the proteins present in the sperm of fertile and subfertile thoroughbred stallions. More specifically, the study focused on stallions that are ‘double-homozygous genotype A/A-A/A’ for SNPs rs397316122 and rs69101140 in exon 5 of the FKBP6 gene, identified as being unique to subfertile horses due to impaired acrosomal exocytosis (IAE), which inhibits the sperm’s ability to fertilize an egg.
  • A comprehensive analysis of the sperm’s proteome – the entirety of proteins expressed – was undertaken. This involved comparing protein abundance in the sperm from frozen/thawed semen samples from both subfertile and fertile stallions.

Significant Findings and Observations

  • Researchers managed to identify a total of 2,220 proteins present in the sperm. Out of these, there were 140 proteins that were differentially abundant, meaning they occurred in significantly different quantities between fertile and subfertile stallions. Specifically, 83 of these proteins were less abundant, and 57 were more abundant in the subfertile stallions compared to their fertile counterparts.
  • It was noted that the proteins that were differentially abundant were predominantly involved in the “metabolism” and “metabolism of lipids” pathways. This suggests that these pathways may have a crucial role in fertility in Thoroughbred stallions.

Specific Findings on Arylsulfatase F and Zona Pellucida Binding

  • A closer examination was made of one protein in particular, arylsulfatase F (ARSF), which was observed through immunofluorescence.
    In sperm from subfertile stallions, a lower proportion of ARSF signals at the acrosome region was observed.
    This may indicate a link between ARSF and fertility in horses.
  • Furthermore, the study employed heterologous zona pellucida binding assays, which revealed that sperm from subfertile stallions tend to bind in smaller proportions to zonae pellucidae (outer membrane of the egg), as compared to sperm from fertile stallions. The decreased binding suggests that sperm from subfertile stallions might face difficulty penetrating the egg’s protective layer.

Conclusion

  • The study concluded that a specific group of proteins, including those of acrosome origin, were found in different amounts in sperm from subfertile stallions with acrosome dysfunction.
  • The differential abundance of these certain proteins and the pathways they play a role in could be pivotal in understanding and possibly addressing fertility issues in Thoroughbred stallions, or other mammals where similar biological processes can be observed.

Cite This Article

APA
Hernández-Avilés C, Ramírez-Agámez L, Weintraub ST, Scoggin CF, Davis BW, Raudsepp T, Varner DD, Love CC. (2024). Proteomic analysis of sperm from fertile stallions and subfertile stallions due to impaired acrosomal exocytosis. Sci Rep, 14(1), 12446. https://doi.org/10.1038/s41598-024-63410-3

Publication

Scientific reports
ISSN: 2045-2322
NlmUniqueID: 101563288
Country: England
Language: English
Volume: 14
Issue: 1
Pages: 12446
PII: 12446

Researcher Affiliations

Hernández-Avilés, Camilo
  • Equine Fertility Laboratory, Department of Large Animal Clinical Sciences, School of Veterinary Medicine and Biomedical Sciences, Texas A&M University, 500 Raymond Stotzer Parkway, College Station, TX, 77843, USA. chernandez@cvm.tamu.edu.
Ramírez-Agámez, Luisa
  • Equine Fertility Laboratory, Department of Large Animal Clinical Sciences, School of Veterinary Medicine and Biomedical Sciences, Texas A&M University, 500 Raymond Stotzer Parkway, College Station, TX, 77843, USA.
Weintraub, Susan T
  • Department of Biochemistry and Structural Biology, The University of Texas Health Science Center, San Antonio, TX, USA.
Scoggin, Charles F
  • LeBlanc Reproduction Center, Rood & Riddle Equine Hospital, Lexington, KY, USA.
Davis, Brian W
  • Veterinary Integrative Biosciences, School of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX, USA.
Raudsepp, Terje
  • Veterinary Integrative Biosciences, School of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX, USA.
Varner, Dickson D
  • Equine Fertility Laboratory, Department of Large Animal Clinical Sciences, School of Veterinary Medicine and Biomedical Sciences, Texas A&M University, 500 Raymond Stotzer Parkway, College Station, TX, 77843, USA.
Love, Charles C
  • Equine Fertility Laboratory, Department of Large Animal Clinical Sciences, School of Veterinary Medicine and Biomedical Sciences, Texas A&M University, 500 Raymond Stotzer Parkway, College Station, TX, 77843, USA.

MeSH Terms

  • Animals
  • Male
  • Horses
  • Proteomics / methods
  • Acrosome Reaction
  • Spermatozoa / metabolism
  • Exocytosis
  • Acrosome / metabolism
  • Infertility, Male / metabolism
  • Infertility, Male / veterinary
  • Infertility, Male / genetics
  • Proteome / metabolism
  • Fertility / genetics
  • Zona Pellucida / metabolism

Conflict of Interest Statement

The authors declare no competing interests.

References

This article includes 98 references
  1. Wassarman PM. Mammalian fertilization: Molecular aspects of gamete adhesion, exocytosis, and fusion.. Cell 1999;96:175–183.
    doi: 10.1016/s0092-8674(00)80558-9pubmed: 9988213google scholar: lookup
  2. Hirohashi N, Yanagimachi R. Sperm acrosome reaction: Its site and role in fertilization.. Biol. Reprod. 2018;99:127–133.
    doi: 10.1093/biolre/ioy045pubmed: 29462288google scholar: lookup
  3. Hernández-Avilés C, Ramírez-Agámez L, Varner DD, Love CC. The stallion sperm acrosome: Considerations from a research and clinical perspective.. Theriogenology 2023;196:121–149.
    doi: 10.1016/j.theriogenology.2022.11.012pubmed: 36413868google scholar: lookup
  4. Varner DD. Subfertility in stallions associated with spermatozoal acrosome dysfunction.. Proc. Am. Assoc. Equine Pract. 2001;47:227–228.
  5. Hernández-Avilés C, Castaneda C, Raudsepp T, Varner DD, Love CC. The role of impaired acrosomal exocytosis in stallion subfertility: A retrospective analysis of the clinical condition, and an update on its diagnosis by high throughput technologies.. Theriogenology 2022;186:40–49.
    doi: 10.1016/j.theriogenology.2022.03.007pubmed: 35429686google scholar: lookup
  6. Hernández-Avilés C, Ramírez-Agámez L, Varner DD, Love CC. Lactate-induced spontaneous acrosomal exocytosis as a method to study acrosomal function in stallions.. Theriogenology 2023;210:169–181.
    doi: 10.1016/j.theriogenology.2023.07.024pubmed: 37517302google scholar: lookup
  7. Raudsepp T. Genome-wide association study implicates testis-sperm-specific FKBP6 as a susceptibility locus for impaired acrosome reactions in stallions.. PLoS Genet. 2012;8:e1003139.
    doi: 10.1371/journal.pgen.1003139pmc: PMC3527208pubmed: 23284302google scholar: lookup
  8. Castaneda C. Thoroughbred stallion fertility is significantly associated with FKBP6 genotype but not with inbreeding or the contribution of a leading sire.. Anim. Genet. 2021;52:813–823.
    doi: 10.1111/age.13142pubmed: 34610162google scholar: lookup
  9. Crackower MA. Essential role of Fkbp6 in male fertility and homologous chromosome pairing in meiosis.. Science 2003;300:1291–1295.
    doi: 10.1126/science.1083022pmc: PMC2882960pubmed: 12764197google scholar: lookup
  10. Miyamato T. Is a genetic defect in Fkbp6 a common cause of azoospermia in humans?. Cell. Mol. Biol. Lett. 2006;11:557–569.
    doi: 10.2478/s11658-006-0043-1pmc: PMC6275806pubmed: 16983454google scholar: lookup
  11. Zhang W, Zhang S, Xiao C, Yang Y, Zhoucum A. Mutation screening of the FKBP6 gene and its association study with spermatogenic impairment in idiopathic infertile men.. Reproduction 2007;133:511–516.
    doi: 10.1530/REP-06-0125pubmed: 17307919google scholar: lookup
  12. Aitken RJ, Nixon B, Lin M, Koppers AJ, Lee YH, Baker MA. Proteomic changes in mammalian spermatozoa during epididymal maturation.. Asian J. Androl. 2007;9:554–564.
    doi: 10.1111/j.1745-7262.2007.00280.xpubmed: 17589795google scholar: lookup
  13. Arcelay E, Salicioni AM, Werthheimer W, Visconti PE. Identification of proteins undergoing tyrosine phosphorylation during mouse sperm capacitation.. Int. J Dev. Biol. 2008;52:463–472.
    doi: 10.1387/ijdb.072555eapubmed: 18649259google scholar: lookup
  14. Amaral A. Identification of proteins involved in human sperm motility using high-throughput differential proteomics.. J. Proteome Res. 2014;13:5670–5684.
    doi: 10.1021/pr500652ypubmed: 25250979google scholar: lookup
  15. Swegen A, Curry BJ, Gibb Z, Lambourne SR, Smith ND, Aitken RJ. Investigation of the stallion sperm proteome by mass spectrometry.. Reproduction 2015;149:235–244.
    doi: 10.1530/REP-14-0500pubmed: 25504869google scholar: lookup
  16. Ortíz-Rodríguez JM. Low glucose and high pyruvate reduce the production of 2-oxoaldehydes, improving mitochondrial efficiency, redox regulation, and stallion sperm function.. Biol. Reprod. 2021;105:519–532.
    doi: 10.1093/biolre/ioab073pubmed: 33864078google scholar: lookup
  17. Kasimanickam RK, Kasimanickam VR, Arangasamy A, Kastelic JP. Sperm and seminal plasma proteomics of high- versus low-fertility Holstein bulls.. Theriogenology 2019;126:41–48.
    doi: 10.1016/j.theriogenology.2018.11.032pubmed: 30529997google scholar: lookup
  18. Mills KM, Aryal UK, Sobreira T, Minton AM, Casey T, Stewart KR. Shotgun proteome analysis of seminal plasma differentiates boars by reproductive performance.. Theriogenology 2020;157:130–139.
    doi: 10.1016/j.theriogenology.2020.07.013pubmed: 32810790google scholar: lookup
  19. Kaya A. Sperm proteins ODF2 and PAWP as markers of fertility in breeding bulls.. Cell Tissue Res. 2022;387:159–171.
    doi: 10.1007/s00441-021-03529-1pubmed: 34762184google scholar: lookup
  20. McReynolds S, Dzieciatkowska M, Stevens J, Hansen KC, Schoolcraft WB, Katz-Jaffe MG. Toward the identification of a subset of unexplained infertility: A sperm proteomic approach.. Fertil. Steril. 2014;102:692–699.
    doi: 10.1016/j.fertnstert.2014.05.021pubmed: 24934493google scholar: lookup
  21. Swegen A, Clulow JR, Baker M, Aitken RJ, Tram QT, Gibb Z. Unraveling infertility: Deciphering the molecular basis of idiopathic infertility in a Thoroughbred stallion.. J. Equine Vet. Sci. 2018;66:90.
    doi: 10.1016/j.jevs.2018.05.056google scholar: lookup
  22. Griffin RA. Proteomic analysis of spermatozoa reveals caseins play a pivotal role in preventing short-term periods of subfertility in stallions.. Biol. Reprod. 2022;106:741–755.
    doi: 10.1093/biolre/ioab225pubmed: 35024820google scholar: lookup
  23. Aebersold R, Mann M. Mass spectrometry-based proteomics.. Nature 2003;422:198–207.
    doi: 10.1038/nature01511pubmed: 12634793google scholar: lookup
  24. Sinha A, Mann M. A beginner’s guide to mass spectrometry-based proteomics.. Biochemist (Lond) 2020;42:64–69.
    doi: 10.1042/BIO20200057google scholar: lookup
  25. Venable JD, Dong MQ, Wohlschlegel J, Dillin A, Yates JR. Automated approach for quantitative analysis of complex peptide mixtures from tandem mass spectra.. Nat. Method. 2004;1:39–45.
    doi: 10.1038/nmeth705pubmed: 15782151google scholar: lookup
  26. Domon B, Aebersold R. Mass spectrometry and protein analysis.. Science 2006;312:212–217.
    doi: 10.1126/science.1124619pubmed: 16614208google scholar: lookup
  27. Nesvizhskii AI. Protein identification by tandem mass spectrometry and sequence database searching.. Methods Mol. Biol. 2007;367:87–119.
    doi: 10.1385/1-59745-575-0:87(2007)pubmed: 17185772google scholar: lookup
  28. Michalski A, Cox J, Mann M. More than 100,000 detectable peptide species elute in a single shotgun prtoemics runs but the majority is inaccessible to data-depended LC-MS/MS.. J. Proteome Res. 2011;10:1785–1793.
    doi: 10.1021/pr101061vpubmed: 21309581google scholar: lookup
  29. Hu A, Noble WS, Wolf-Yadlin A. Technical advances in proteomics: New developments in data-independent acquisition.. F1000Research 2016;5:419.
    doi: 10.12688/f100research.7042.1.eCollection2016pmc: PMC4821292pubmed: 27092249google scholar: lookup
  30. Gillet LC. Targeted data extraction of the MS/MS spectra generated by data-independent acquisition: A new concept for consistent and accurate proteome analysis.. Mol. Cell Proteom. 2012;11(O111):016717.
    doi: 10.1074/mcp.O111.01717pmc: PMC3433915pubmed: 22261725google scholar: lookup
  31. Law KP, Lim YP. Recent advances in mass spectrometry: Data independent analysis and hyper reaction monitoring.. Expert Rev. Proteom. 2013;10:551–566.
    doi: 10.1586/14789450.2013.858022pubmed: 24206228google scholar: lookup
  32. Ludwig C, Gillet L, Rosenberger G, Amon S, Collins BC, Aebersold R. Data-independent acquisition-based SWATH-MS for quantitative proteomics: A tutorial.. Mol. Syst. Biol. 2018;14:e8126.
    doi: 10.15252/msb.20178126pmc: PMC6088389pubmed: 30104418google scholar: lookup
  33. Zhang W, Ge W, Ruan G, Cai X, Guo T. Data-independent acquisition mass spectrometry-based proteomics and software tools: A glimpse in 2020.. Proteomics 2020;10:e1900276.
    doi: 10.1002/pmic201900276pubmed: 32275110google scholar: lookup
  34. Secciani F. Protein profile of capacitated versus ejaculated human sperm.. J. Proteome Res. 2009;8:3377–3389.
    doi: 10.1021/pr900031rpubmed: 19408963google scholar: lookup
  35. Castillo J. Proteomic changes in human sperm during sequential in vitro capacitation and acrosome reaction.. Front. Cell. Dev. Biol. 2019;7:295.
    doi: 10.3389/fcell.2019.00295pmc: PMC6879431pubmed: 31824947google scholar: lookup
  36. Novak S. Biomarkers of in vivo fertility in sperm and seminal plasma of fertile stallions.. Theriogenology 2010;74:956–967.
    doi: 10.1016/j.theriogenology.2010.04.025pubmed: 20580075google scholar: lookup
  37. Griffin RA. Mass spectrometry reveals distinct proteomic profiles in high- and low-quality stallion spermatozoa.. Reproduction 2020;160:695–707.
    doi: 10.1530/REP-20-0284pubmed: 32805711google scholar: lookup
  38. Martín-Cano FE. Proteomic profilling of stallion spermatozoa suggests changes in sperm metabolism and compromised redox regulation after cryopreservation.. J. Proteom. 2020;221:103765.
    doi: 10.1016/j.jprot.2020.103765pubmed: 32247875google scholar: lookup
  39. Gaitskell-Phillips G. Seminal plasma AnnexinA2 protein is a relevant biomarker for stallions which require removal of seminal plasma for sperm survival upon refrigeration.. Biol. Reprod. 2020;103:1275–1288.
    doi: 10.1093/biolre/ioaa153pubmed: 32857155google scholar: lookup
  40. Gaitskell-Phillips G. Differences in the proteome of stallion spermatozoa explain stallion-to-stallion variability in sperm quality post-thaw.. Biol. Reprod. 2021;104:1097–1113.
    doi: 10.1093/biolre/ioab003pubmed: 33438027google scholar: lookup
  41. Gaitskell-Phillips G. In stallion spermatozoa, superoxide dismutase (Cu-Zn) (SOD) and the aldo-keto-reductase family 1 member b (AKR1B1) are the proteins most significantly reduced by cryopreservation.. J. Proteome. Res. 2021;20:2435–2446.
    doi: 10.1021/acs.jproteome.0c00932pmc: PMC8562871pubmed: 33656888google scholar: lookup
  42. Langlais J, Kan FW, Granger L, Raymond L, Bleau G, Roberts KD. Identification of sterol acceptors that stimulate cholesterol efflux from human spermatozoa during in vitro capacitation.. Gamete Res. 1988;20:185–201.
    doi: 10.1002/mrd.1120200209pubmed: 3235036google scholar: lookup
  43. Osheroff JE, Visconti PE, Valenzuela JP, Travis AJ, Alvarez J, Kopf GS. Regulation of human sperm capacitation by a cholesterol efflux-stimulated signal transduction pathway leading to protein kinase A-mediated up-regulation of protein tyrosine phosphorylation.. Mol. Hum. Reprod. 1999;5:1017–1026.
    doi: 10.1093/molehr/5.11.1017pubmed: 10541563google scholar: lookup
  44. Visconti PE. Cholesterol efflux-mediated signal transduction in mammalian sperm. Beta-cyclodextrins initiate transmembrane signaling leading to an increase in protein tyrosine phosphorylation and capacitation.. J. Biol. Chem. 1999;274:3235–3242.
    doi: 10.1074/jbc.274.5.3235pubmed: 9915865google scholar: lookup
  45. Visconti PE. Cholesterol efflux-mediated signal transduction in mammalian sperm: cholesterol release signals an increase in protein tyrosine phosphorylation during mouse sperm capacitation.. Dev. Biol. 1999;214:429–443.
    doi: 10.1006/dbio/1999/9428pubmed: 10525345google scholar: lookup
  46. Flesch FM. Bicarbonate stimulated phospholipid scrambling induces cholesterol redistribution and enables cholesterol depletion in the sperm plasma membrane.. J. Cell Sci. 2001;114:3543–3555.
    doi: 10.1242/jcs.114.19.3543pubmed: 11682613google scholar: lookup
  47. Breitbart H, Lax X, Rotem R, Naor Z. Role of protein kinase C in the acrosome reaction of mammalian spermatozoa.. Biochem. J. 1992;281:473–476.
    doi: 10.1042/bj2810473pmc: PMC1130709pubmed: 1736894google scholar: lookup
  48. Spungin B, Breitbart H. Calcium mobilization and influx during sperm exocytosis.. J. Cell Sci. 1996;109:1947–1955.
    doi: 10.1242/jcs.109.7.1947pubmed: 8832417google scholar: lookup
  49. Liu DY, Baker HW. Protein kinase C plays an important role in the human zona pellucida-induced acrosome reaction.. Mol. Hum. Reprod. 1997;12:1037–1043.
    doi: 10.1093/molehr/3.12.1037pubmed: 9464848google scholar: lookup
  50. Branham MT, Mayorga LS, Tomes CN. Calcium-induced acrosomal exocytosis requires cAMP acting through a protein kinase A-independent, Epac-mediated pathway.. J. Biol. Chem. 2006;281:8656–8666.
    doi: 10.1074/jbc.M508854200pubmed: 16407249google scholar: lookup
  51. Singleton CL, Killian GJ. A study of phospholipase in albumin and its role in inducing the acrosome reaction of guinea pig spermatozoa in vitro.. J. Androl. 1983;4:150–156.
    doi: 10.1002/j.1939-4640.1983.tb00740.xpubmed: 6853360google scholar: lookup
  52. Flemming AD, Yanagimachi R. Evidence suggesting the importance of fatty acids and the fatty acid moieties of sperm membrane phospholipids in the acrosome reaction of guinea pig spermatozoa.. J. Exp. Zool. 1984;229:485–489.
    doi: 10.1002/jez.1402290317pubmed: 6423766google scholar: lookup
  53. Lepage N, Roberts KD. Purification of lysophospholipase of human spermatozoa and its implication in the acrosome reaction.. Biol. Reprod. 1995;52:616–624.
    doi: 10.1095/biolreprod52.3.616pubmed: 7756455google scholar: lookup
  54. Dominguez L, Yunes RM, Fornés MW, Burgos M, Mayorga LS. Calcium and phospholipase A2 are both required for the acrosome reaction mediated by G-proteins simulation in human spermatozoa.. Mol. Reprod. Dev. 1999;52:297–302.
    doi: 10.1002/(SICI)1098-2795(199903)52:3<297::AID-MRD7>3.0.CO;2-Tpubmed: 10206661google scholar: lookup
  55. Li K, Jin JY, Chen WY, Shi QX, Ni Y, Roldan ERS. Secretory phospholipase A2 group IID is involved in progesterone-induced acrosomal exocytosis of human spermatozoa.. J. Androl. 2012;33:975–983.
    doi: 10.2164/jandrol.111.014886pubmed: 22240557google scholar: lookup
  56. de Lamirande E, Tsai C, Harakat A, Gagnon C. Involvement of reactive oxygen species in human sperm acrosome reaction induced by A23187, lysophosphatidylcholine, and biological fluid ultrafiltrates.. J. Androl. 1998;19:585–594.
    doi: 10.1002/j.1939-4640/1998.tb02061.xpubmed: 9796619google scholar: lookup
  57. O’Flaherty C, Beorlegui NB, Beconi MT. Reactive oxygen species requirements for bovine sperm capacitation and acrosome reaction.. Theriogenology 1999;52:289–301.
    doi: 10.1016/S0093-691X(99)00129-6pubmed: 10734395google scholar: lookup
  58. Aitken RJ. Reactive oxygen species as mediators of sperm capacitation and pathological damage.. Mol. Reprod. Dev. 2017;84:1039–1052.
    doi: 10.1002/mrd.22871pubmed: 28749007google scholar: lookup
  59. Brinsko SP, Love CC, Bauer JE, Macpherson ML, Varner DD. Cholesterol-to-phospholipid ratio in whole sperm and seminal plasma from fertile stallions and stallions with unexplained subfertility.. Anim. Reprod. Sci. 2007;99:65–71.
    doi: 10.1016/j.anireprosci.2006.03.018pubmed: 16713689google scholar: lookup
  60. Conrad DJ. The arachidonate 12/15 lipoxygenases. A review of tissue expression and biologic function.. Clin. Rev. Allergy Immunol. 1999;17:71–89.
    doi: 10.1007/BF02737598pubmed: 10436860google scholar: lookup
  61. Meizel S, Turner KO. The effects of products and inhibitors of arachidonic acid metabolism on the hamster sperm acrosome reaction.. J. Exp. Zool. 1984;231:283–288.
    doi: 10.1002/jez.1402310213pubmed: 6434693google scholar: lookup
  62. Lax Y, Grossman S, Rubinstein S, Magid N, Breitbart H. Role of lipoxygenase in the mechanism of acrosome reaction in mammalian spermatozoa.. Biochim. Biophys. Acta. 1990;1043:12–18.
    doi: 10.1016/0005-2760(90)90104-6pubmed: 2106918google scholar: lookup
  63. Mack SR, Han HL, De Jonge CJ, Anderson RA, Zaneveld LJD. The human sperm acrosome reaction does not depend on arachidonic acid metabolism via the cyclooxygenase and lipoxygenase pathways.. J. Androl. 1992;13:551–559.
    doi: 10.1002/j.1939-4640.1992.tb00351.xpubmed: 1338069google scholar: lookup
  64. Walters JLH. Pharmacological inhibition of arachidonate 15-lipoxygenase protects human spermatozoa against oxidative stress.. Biol. Reprod. 2018;98:784–794.
    doi: 10.1093/biolre/ioy058pubmed: 29546268google scholar: lookup
  65. Nimmo MR, Cross NL. Structural features of sterols required to inhibit human sperm capacitation.. Biol. Reprod. 2003;68:1308–1317.
    doi: 10.1095/biolreprod.102.008607pubmed: 12606419google scholar: lookup
  66. Veitia RA, Ottolenghi C, Bissery MC, Fellous A. A novel human gene, encoding a potential membrane protein conserved from yeast to man, is strongly expressed in testis and cancer cell lines.. Cytogenet. Cell. Genet. 1999;85:217–220.
    doi: 10.1159/000015296pubmed: 10449901google scholar: lookup
  67. Mo C, Vlachovic M, Bard M. The ERG28-encoded protein, Erg28p, interacts with both the sterol C-4 demethylation enzyme complex as well as the late biosynthetic protein, the C-24 sterol methyltransferase (Erg6p). Biochim. Biophys. Acta. 2004;1686:30–36.
    doi: 10.1016/j.bbalip.2004.08.001pubmed: 15522820google scholar: lookup
  68. Zeng Y, Clark EN, Florman HM. Sperm membrane potential: Hyperpolarization during capacitation regulates zona pellucida-dependent acrosomal secretion.. Dev. Biol. 1995;171:554–563.
    doi: 10.1006/dbio.1995.1304pubmed: 7556936google scholar: lookup
  69. Garcia MA, Meizel S. Regulation of intracellular pH in capacitated human spermatozoa by a Na+/H+ exchanger.. Mol. Reprod. Dev. 1999;52:189–195.
    doi: 10.1002/(SICI)1098-2795(199902)52:2<189::AID-MRD10>3.0.CO;2-Dpubmed: 9890750google scholar: lookup
  70. Nikolajczyk BS, Rand MG. Characterization of rabbit testis β-galactosidase and arylsulfatase A: Purification and localization in spermatozoa during the acrosome reaction.. Biol. Reprod. 1992;46:366–378.
    doi: 10.1095/biolreprod46.3.366pubmed: 1352147google scholar: lookup
  71. Tantibhedhyangkul J. Role of sperm surface arylsulfatase A in mouse sperm-zona pellucida binding.. Biol. Reprod. 2002;67:212–219.
    doi: 10.1095/biolreprod67.1.212pubmed: 12080020google scholar: lookup
  72. Gomez-Torres MJ. Arylsulfatase A remodeling during human sperm in vitro capacitation using field emission scanning electron microscopy (FE-SEM). Cells 2021;10:222.
    doi: 10.3390/cells10020222pmc: PMC7912702pubmed: 33498624google scholar: lookup
  73. Carmona E. Arylsulfatase A is present on the pig sperm surface and is involved in sperm-zona pellucida binding.. Dev. Biol. 2002;247:182–196.
    doi: 10.1006/dbio.2002.0690pubmed: 12074561google scholar: lookup
  74. Carmona E. Binding of arylsulfatase A to mouse sperm inhibits gamete interaction and induces the acrosome reaction.. Biol. Reprod. 2002;66:1820–1827.
    doi: 10.1095/biolreprod66.6.1820pubmed: 12021068google scholar: lookup
  75. Xu H. Sperm arylsulfatase A binds to mZP2 and mZP3 glycoproteins in a nonenzymatic manner.. Reproduction 2012;144:209–219.
    doi: 10.1530/REP-11-0338pubmed: 22685254google scholar: lookup
  76. Lin YN, Roy A, Yan W, Burns KH, Matzuk MM. Loss of zona pellucida binding proteins in the acrosomal matrix disrupts acrosome biogenesis and sperm morphogenesis.. Mol. Cell. Biol. 2007;27:6794–6805.
    doi: 10.1128/MCB.01029-07pmc: PMC2099232pubmed: 17664285google scholar: lookup
  77. Richardson RT, Yamasaki N, O’Rand MG. Sequence of a rabbit sperm zona pellucida binding protein and localization during the acrosome reaction.. Dev. Biol. 1994;165:688–701.
    doi: 10.1006/dbio.1994.1285pubmed: 7525387google scholar: lookup
  78. Swegen A, Smith ND, Gibb Z, Curry BJ, Aitken RJ. The serine protease testisin is present on the surface of capacitated stallion spermatozoa and interacts with key zona pellucida binding proteins.. Andrology 2019;7:199–212.
    doi: 10.1111/andr.12569pubmed: 30549223google scholar: lookup
  79. Ramírez-Agámez L, Hernández-Avilés C, Ortíz I, Love CC, Varner DD, Hinrichs K. Lactate as the sole energy substrate induces spontaneous acrosome reaction in viable stallion sperm.. Andrology 2024;12:459–471.
    doi: 10.1111/andr.13479pubmed: 37300872google scholar: lookup
  80. Commerford KL, Love CC, Brinsko SP, Waite JA, Teague SR, Varner DD. Validation of a commercially available fluorescence-based instrument to evaluate stallion spermatozoal concentration.. Anim. Reprod. Sci. 2008;107:314.
    doi: 10.1016/j.anireprosci.2008.05.093google scholar: lookup
  81. Varner DD, Vaughan SD, Johnson L. Use of a computerized system for evaluation of equine spermatozoal motility.. Am. J. Vet. Res. 1991;52:224–230.
    doi: 10.2460/ajvr.1991.52.02.224pubmed: 2012333google scholar: lookup
  82. Kenney RM, Hurtgen J, Pierson R, Witherspoon D, Simons J. Society for Theriogenology Manual for Clinical Fertility Evaluation of the Stallion.. Hastings, Nebraska. p. 100. (1983).
  83. Love CC, Kenney RM. The relationship of increased susceptibility of sperm DNA to denaturation and fertility in the stallion.. Theriogenology 1998;50:955–972.
    doi: 10.1016/S0093-691X(98)00199-Xpubmed: 10734467google scholar: lookup
  84. Waite JA. Factors impacting equine sperm recovery rate and quality following cushioned centrifugation.. Theriogenology 2008;70:704–714.
    doi: 10.1016/j.theriogenology.2008.04.04pubmed: 18573520google scholar: lookup
  85. Hernández-Avilés C, Ramírez-Agámez L, Varner DD, Love CC. Effects of egg yolk level, penetrating cryoprotectant, and pre-freeze cooling rate, on the post-thaw quality of stallion sperm.. Anim Reprod Sci. 2023;248:107162.
    doi: 10.1016/j.anireprosci.2022.107162pubmed: 36469980google scholar: lookup
  86. Love CC, Merlo B, Rizzato G, Mislei B, Castegnetti C, Mari G. Recovery rate and sperm quality after centrifugation of stallion sperm in different gradient concentrations.. J. Equine Vet. Sci. 2014;34:63.
    doi: 10.1016/j.jevs.2013.10.038google scholar: lookup
  87. Ortíz I, Félix M, Resènde H, Ramírez-Agámez L, Love CC, Hinrichs K. Flow-cytometric analysis of membrane integrity of stallion sperm in the face of agglutination: The “zombie sperm” dilemma.. J. Assist. Reprod. Gen. 2021;38:2465–2480.
    doi: 10.1007/s10815-021-02134-zpmc: PMC8490572pubmed: 33991296google scholar: lookup
  88. Perfetto SP. Amine reactive dyes: An effective tool to discriminate live and dead cells in polychromatic flow cytometry.. J. Immunol. Methods. 2006;313:199–208.
    doi: 10.1016/j.jim.2006.04.007pubmed: 16756987google scholar: lookup
  89. Cross NL, Morales P, Overstreet JW, Hanson FW. Two simple methods for detecting acrosome-reacted human sperm.. Gamete Res. 1986;15:213–226.
    doi: 10.1002/mrd.1120150303google scholar: lookup
  90. Searle BC. Chromatogram libraries improve peptide detection and quantification by data-independent acquisition mass spectrometry.. Nat. Commun. 2018;9:5128.
    doi: 10.1038/s41467-018-07454-wpmc: PMC6277451pubmed: 30510204google scholar: lookup
  91. Gessulat S. Prosit: Proteome-wide prediction of peptide tandem mass spectra by deep learning.. Nat. Methods. 2019;16:509–518.
    doi: 10.1038/s41292-019-0426-7pubmed: 31133760google scholar: lookup
  92. Chambers MC. A cross-platform toolkit for mass spectrometry and proteomics.. Nat. Biotechnol. 2012;30:918–920.
    doi: 10.1038/nbt.2377pmc: PMC3471674pubmed: 23051804google scholar: lookup
  93. Käll L, Storey JD, Noble WS. Non-parametric estimation of posterior error probabilities associated with peptides identified by tandem mass spectrometry.. Bioinformatics 2008;24:42–48.
    doi: 10.1093/bioinformatics/btn294pmc: PMC2732210pubmed: 18689838google scholar: lookup
  94. Benjamini Y, Hochberg Y. Controlling the false discovery rate: A practical and powerful approach to multiple testing.. J. Royal Stat. Soc. Ser. B. 1995;57:125–133.
    doi: 10.1111/j.2517-6161.1995.tb02031.xgoogle scholar: lookup
  95. Raudvere U. g:Profiler: A web server for functional enrichment analysis and conversions of gene lists.. Nucleic Acids Res. 2019;47:191–198.
    doi: 10.1093/nar/gkz369pmc: PMC6602461pubmed: 31066453google scholar: lookup
  96. Mlecnik B, Galon J, Bindea G. Comprehensive functional analysis of large lists of genes and proteins.. J. Proteom. 2018;171:2–10.
    doi: 10.1016/j.jprot.2017.03.016pubmed: 28343001google scholar: lookup
  97. Mugnier S, Dell’Aquila ME, Pelaez J, Douet C, Ambruosi B, De Santis T, Lacalandra GM, Lebos C, Sizaret PY, Delaleu B, Monget P, Mermillod P. New insights into the mechanisms of fertilization: Comparison of the fertilization steps, composition, and structure of the zona pellucida between horses and pigs.. Biol. Reprod. 2009;81:856–870.
    doi: 10.1095/biolreprod.109.077651pubmed: 19587333google scholar: lookup
  98. Sessions-Bresnahan D, Graham JK, Carnevale EM. Validation of a heterologous fertilization assay and comparison of fertilization rates of equine oocytes using in vitro fertilization, perivitelline, and intracytoplasmic sperm injections.. Theriogenology 2014;82:274–282.
    doi: 10.1016/j.theriogenology.2014.04.002(2014)pubmed: 24815920google scholar: lookup

Citations

This article has been cited 0 times.
Subscribe to our newsletter
Join 99,344 horse owners like you who receive our email updates on horse health and nutrition.