Oviduct binding and elevated environmental ph induce protein tyrosine phosphorylation in stallion spermatozoa.
Abstract: Sperm-oviduct binding is an essential step in the capacitation process preparing the sperm for fertilization in several mammalian species. In many species, capacitation can be induced in vitro by exposing spermatozoa to bicarbonate, Ca(2+), and albumin; however, these conditions are insufficient in the horse. We hypothesized that binding to the oviduct epithelium is an essential requirement for the induction of capacitation in stallion spermatozoa. Sperm-oviduct binding was established by coincubating equine oviduct explants for 2 h with stallion spermatozoa (2 × 10(6) spermatozoa/ml), during which it transpired that the highest density (per mm(2)) of oviduct-bound spermatozoa was achieved under noncapacitating conditions. In subsequent experiments, sperm-oviduct incubations were performed for 6 h under noncapacitating versus capacitating conditions. The oviduct-bound spermatozoa showed a time-dependent protein tyrosine phosphorylation response, which was not observed in unbound spermatozoa or spermatozoa incubated in oviduct explant conditioned medium. Both oviduct-bound and unbound sperm remained motile with intact plasma membrane and acrosome. Since protein tyrosine phosphorylation can be induced in equine spermatozoa by media with high pH, the intracellular pH (pHi) of oviduct explant cells and bound spermatozoa was monitored fluorometrically after staining with BCECF-AM dye. The epithelial secretory cells contained large, alkaline vesicles. Moreover, oviduct-bound spermatozoa showed a gradual increase in pHi, presumably due to an alkaline local microenvironment created by the secretory epithelial cells, given that unbound spermatozoa did not show pHi changes. Thus, sperm-oviduct interaction appears to facilitate equine sperm capacitation by creating an alkaline local environment that triggers intracellular protein tyrosine phosphorylation in bound sperm.
© 2014 by the Society for the Study of Reproduction, Inc.
Publication Date: 2014-05-14 PubMed ID: 24829033DOI: 10.1095/biolreprod.113.116418Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research article discusses the process involved in preparing stallion sperm for fertilization, specifically the role of binding to oviduct epithelium and the impact of elevated environmental pH. It is suggested that this interaction aids in sperm capacitation by promoting an alkaline environment, leading to protein tyrosine phosphorylation within the sperm.
Research Objectives and Hypothesis
- The primary objective of this research was to understand the role of sperm-oviduct binding in inducing capacitation – a process that prepares sperm for fertilization in horses.
- The researchers hypothesized that binding of stallion spermatozoa to the oviduct epithelium is an essential requirement for capacitation.
Research Methods
- Experiments were conducted by coincubating equine oviduct explants with stallion spermatozoa for 2 hours. The density of oviduct-bound sperm was observed under noncapacitating conditions.
- Further, they performed sperm-oviduct incubations for 6 hours under capacitating and noncapacitating conditions.
- The researchers monitored the protein tyrosine phosphorylation response of the oviduct-bound sperm over time.
- They also monitored the intracellular pH of the oviduct explant cells and bound sperm using BCECF-AM dye.
Findings
- It was observed that the oviduct-bound sperm showed a time-dependent protein tyrosine phosphorylation response, an occurrence not noticed in unbound sperm.
- The researchers found that both oviduct-bound and unbound sperm remained motile, with intact plasma membrane and acrosome.
- Secretory cells of the oviduct were found to contain large, alkaline vesicles. Oviduct-bound sperm showed a gradual increase in intracellular pH, hypothesized to possibly be due to an alkaline local environment created by secretory epithelial cells.
Conclusion
- Based on these findings, it appears that the interaction between sperm and the oviduct may facilitate equine sperm capacitation. This is potentially due to the alkaline local microenvironment, which triggers protein tyrosine phosphorylation in the sperm.
Cite This Article
APA
Leemans B, Gadella BM, Sostaric E, Nelis H, Stout TA, Hoogewijs M, Van Soom A.
(2014).
Oviduct binding and elevated environmental ph induce protein tyrosine phosphorylation in stallion spermatozoa.
Biol Reprod, 91(1), 13.
https://doi.org/10.1095/biolreprod.113.116418 Publication
Researcher Affiliations
- Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Ghent, Belgium.
- Departments of Farm Animal Health, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands Biochemistry and Cell Biology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands b.m.gadella@uu.nl.
- Equine Sciences, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
- Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Ghent, Belgium.
- Departments of Farm Animal Health, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands Equine Sciences, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
- Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Ghent, Belgium.
- Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Ghent, Belgium.
MeSH Terms
- Animals
- Female
- Fertilization / physiology
- Horses
- Male
- Oviducts / metabolism
- Phosphorylation
- Protein-Tyrosine Kinases / metabolism
- Sperm Capacitation / physiology
- Spermatozoa / metabolism
- Tyrosine / metabolism
Citations
This article has been cited 9 times.- Felix MR, Turner RM, Dobbie T, Hinrichs K. Successful in vitro fertilization in the horse: production of blastocysts and birth of foals after prolonged sperm incubation for capacitation†. Biol Reprod 2022 Dec 10;107(6):1551-1564.
- Leemans B, Bromfield EG, Stout TAE, Vos M, Van Der Ham H, Van Beek R, Van Soom A, Gadella BM, Henning H. Developing a reproducible protocol for culturing functional confluent monolayers of differentiated equine oviduct epithelial cells†. Biol Reprod 2022 Apr 26;106(4):710-729.
- Mahé C, Zlotkowska AM, Reynaud K, Tsikis G, Mermillod P, Druart X, Schoen J, Saint-Dizier M. Sperm migration, selection, survival, and fertilizing ability in the mammalian oviduct†. Biol Reprod 2021 Aug 3;105(2):317-331.
- Seifi-Jamadi A, Zhandi M, Kohram H, Luceño NL, Leemans B, Henrotte E, Latour C, Demeyere K, Meyer E, Van Soom A. Influence of seasonal differences on semen quality and subsequent embryo development of Belgian Blue bulls. Theriogenology 2020 Dec;158:8-17.
- Leemans B, Stout TAE, Soom AV, Gadella BM. pH-dependent effects of procaine on equine gamete activation†. Biol Reprod 2019 Nov 21;101(5):1056-1074.
- Kuo YW, Li SH, Maeda K, Gadella BM, Tsai PS. Roles of the reproductive tract in modifications of the sperm membrane surface. J Reprod Dev 2016 Aug 25;62(4):337-43.
- Saint-Dizier M, Souza-Fabjan JMG, Reynaud K, Mermillod P, Almiñana C, Bauersachs S, Mahé C. Oviduct epithelium interactions: roles in sperm selection and embryo quality. Anim Reprod 2025;22(3):e20250035.
- Leemans B, Gadella BM, Marchand JHEAM, Van Soom A, Stout TAE. Induction of in vivo-like ciliation in confluent monolayers of re-differentiated equine oviduct epithelial cells†. Biol Reprod 2024 Sep 14;111(3):580-599.
- Sushadi PS, Kuwabara M, Maung EEW, Mohamad Mohtar MS, Sakamoto K, Selvaraj V, Asano A. Arresting calcium-regulated sperm metabolic dynamics enables prolonged fertility in poultry liquid semen storage. Sci Rep 2023 Dec 8;13(1):21775.
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