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Home / Equine Research Bank / Andrology / Effect of bicarbonate and polyvinyl alcohol on in vitro capa...
Andrology2024; 13(2); 382-395; doi: 10.1111/andr.13667

Effect of bicarbonate and polyvinyl alcohol on in vitro capacitation and fertilization ability of cryopreserved equine spermatozoa.

Abstract: Factors contributing to the limited success of in vitro fertilization in horses remain to be studied. In this work, we elucidated the effect of different essential capacitation media components, bicarbonate, and bovine serum albumin or polyvinyl-alcohol, and the incubation microenvironment on sperm parameters associated with capacitation, acrosome reaction, and their ability to activate oocytes via heterologous intracytoplasmic spermatozoa injection in equine cryopreserved spermatozoa. Methods: Frozen-thawed spermatozoa underwent incubation at different time intervals in either Tyrode's albumin lactate pyruvate medium (non-capacitating; NC) or Tyrode's albumin lactate pyruvate supplemented with bicarbonate, bicarbonate and polyvinyl-alcohol, bicarbonate and bovine serum albumin, polyvinyl-alcohol and bovine serum albumin alone. Protein kinase A-phosphorylated substrates and tyrosine phosphorylation levels, sperm motility, and acrosome reaction percentages were evaluated. After determining the best condition media (capacitating; CAP), heterologous intracytoplasmic spermatozoa injection on pig oocytes was performed and the phospholipase C zeta sperm localization pattern was evaluated. Results: Incubation of frozen-thawed equine spermatozoa with bicarbonate and polyvinyl-alcohol in atmospheric air for 45 min induced an increase in protein kinase A-phosphorylated substrates and tyrosine phosphorylation levels compared to NC condition. Sperm incubation in bicarbonate and polyvinyl-alcohol medium showed an increase in total motility and progressive motility with respect to NC (p ≤ 0.05). Interestingly, three parameters associated with sperm hyperactivation were modulated under bicarbonate and polyvinyl-alcohol conditions. The kinematic parameters curvilinear velocity and amplitude of lateral head displacement significantly increased, while straightness significantly diminished (curvilinear velocity: bicarbonate and polyvinyl-alcohol = 120.9 ± 2.9 vs. NC = 76.91 ± 6.9 µm/s) (amplitude of lateral head displacement: bicarbonate and polyvinyl-alcohol = 1.15 ± 0.02 vs. NC = 0.77 ± 0.03 µm) (straightness: bicarbonate and polyvinyl-alcohol = 0.76 ± 0.01 vs. NC = 0.87 ± 0.02) (p ≤ 0.05). Moreover, the spontaneous acrosome reaction significantly increased in spermatozoa incubated in this condition. Finally, bicarbonate and polyvinyl-alcohol medium was established as CAP medium. Although no differences were found in phospholipase C zeta localization pattern in spermatozoa incubated under CAP, equine spermatozoa pre-incubated in CAP condition for 45 min showed higher fertilization rates when injected into matured pig oocytes (NC: 47.6% vs. CAP 76.5%; p ≤ 0.05). Conclusions: These findings underscore the importance of bicarbonate and polyvinyl-alcohol in supporting critical events associated with in vitro sperm capacitation in the horse, resulting in higher oocyte activation percentages following heterologous intracytoplasmic spermatozoa injection. This protocol could have an impact on reproductive efficiency in the equine breeding industry.
© 2024 American Society of Andrology and European Academy of Andrology.
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Publication Date: 2024-05-28 PubMed ID: 38804843DOI: 10.1111/andr.13667Google Scholar: Lookup
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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 explores the impact of bicarbonate and polyvinyl alcohol on in vitro capacitation and fertilization effectiveness of frost-protected horse spermatozoa. The research findings highlight the beneficial role of bicarbonate and polyvinyl alcohol in enhancing crucial events related to in vitro sperm capacitation in horses, resulting in increased oocyte stimulation rates after heterologous intracytoplasmic sperm injection.

Research Methodology

  • The researchers used frozen-thawed horse spermatozoa for the study.
  • The spermatozoa were incubated in different conditions with variation in levels of bicarbonate and two proteins: polyvinyl-alcohol and bovine serum albumin.
  • The team evaluated the spermatozoa for Protein kinase A (PKA)-phosphorylated substrates and tyrosine phosphorylation levels, sperm motility, and acrosome reaction percentages.
  • Further, heterologous intracytoplasmic spermatozoa injection was performed on pig oocytes using the best condition media (capacitating; CAP). The phospholipase C zeta sperm localization pattern was then evaluated.

Research Findings

  • A significant increase in PKA-phosphorylated substrates and tyrosine phosphorylation levels were observed when the spermatozoa were incubated with bicarbonate and polyvinyl-alcohol in atmospheric air for 45 minutes.
  • An uptick in total motility and progressive motility of the sperm was also seen with incubation in bicarbonate and polyvinyl-alcohol medium.
  • Three parameters associated with sperm hyperactivation witnessed considerable modulation under bicarbonate and polyvinyl-alcohol conditions. Curvilinear velocity and amplitude of lateral head displacement increased significantly. However, straightness decreased.
  • Incubation in bicarbonate and polyvinyl-alcohol medium resulted in an increase in spontaneous acrosome reaction in the spermatozoa.
  • There was an apparent increase in the fertilization rates when the spermatozoa pre-incubated in this established CAP condition were injected into matured pig oocytes compared to Non-Capacitating (NC) condition.

Conclusions

  • The study points out the importance of bicarbonate and polyvinyl-alcohol in supporting critical events associated with in vitro sperm capacitation in horses.
  • Usage of these components resulted in higher oocyte activation percentages following heterologous intracytoplasmic spermatozoa injection.
  • The implications of this study may prove highly significant in enhancing reproductive efficiency in the equine breeding industry.

Cite This Article

APA
Arroyo-Salvo C, Río S, Bogetti ME, Plaza J, Miragaya M, Yaneff A, Davio C, Fissore R, Gervasi MG, Gambini A, Perez-Martinez S. (2024). Effect of bicarbonate and polyvinyl alcohol on in vitro capacitation and fertilization ability of cryopreserved equine spermatozoa. Andrology, 13(2), 382-395. https://doi.org/10.1111/andr.13667

Publication

Andrology
ISSN: 2047-2927
NlmUniqueID: 101585129
Country: England
Language: English
Volume: 13
Issue: 2
Pages: 382-395

Researcher Affiliations

Arroyo-Salvo, Camila
  • Centro de Estudios Farmacológicos y Botánicos (CEFYBO), CONICET-UBA, Buenos Aires, Argentina.
Río, Sofía
  • Centro de Estudios Farmacológicos y Botánicos (CEFYBO), CONICET-UBA, Buenos Aires, Argentina.
Bogetti, María Eugenia
  • Centro de Estudios Farmacológicos y Botánicos (CEFYBO), CONICET-UBA, Buenos Aires, Argentina.
Plaza, Jessica
  • Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, INITRA, Buenos Aires, Argentina.
Miragaya, Marcelo
  • Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, INITRA, Buenos Aires, Argentina.
Yaneff, Agustín
  • Instituto de Investigaciones Farmacológicas (ININFA-UBA-CONICET), Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires, Argentina.
Davio, Carlos
  • Instituto de Investigaciones Farmacológicas (ININFA-UBA-CONICET), Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires, Argentina.
Fissore, Rafael
  • Department of Veterinary and Animal Sciences, University of Massachusetts, Amherst, Massachusetts, USA.
Gervasi, María Gracia
  • Department of Veterinary and Animal Sciences, University of Massachusetts, Amherst, Massachusetts, USA.
  • Department of Animal Science, University of Connecticut, Storrs, Connecticut, USA.
Gambini, Andrés
  • School of Agriculture and Food Sustainability, The University of Queensland, Gatton, Queensland, Australia.
Perez-Martinez, Silvina
  • Centro de Estudios Farmacológicos y Botánicos (CEFYBO), CONICET-UBA, Buenos Aires, Argentina.

MeSH Terms

  • Animals
  • Male
  • Horses
  • Cryopreservation / veterinary
  • Sperm Capacitation / drug effects
  • Bicarbonates / pharmacology
  • Semen Preservation / veterinary
  • Semen Preservation / methods
  • Spermatozoa / drug effects
  • Spermatozoa / physiology
  • Polyvinyl Alcohol / pharmacology
  • Sperm Motility / drug effects
  • Female
  • Acrosome Reaction / drug effects
  • Sperm Injections, Intracytoplasmic / veterinary
  • Fertilization in Vitro / veterinary

Grant Funding

  • PICT 2018 01681 / ANPCYT (FONCyT, Argentina-Préstamo BID)
  • PICT 2021-I-A-00129 / ANPCYT (FONCyT, Argentina-Préstamo BID)
  • 2022-67016-36302 / National Institute of Food and Agriculture

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