An alkaline follicular fluid fraction induces capacitation and limited release of oviduct epithelium-bound stallion sperm.
Abstract: Induction of hyperactivated motility is considered essential for triggering the release of oviduct-bound mammalian spermatozoa in preparation for fertilization. In this study, oviduct-bound stallion spermatozoa were exposed for 2 h to: i) pre-ovulatory and ii) post-ovulatory oviductal fluid; iii) 100% and iv) 10% follicular fluid (FF); v) cumulus cells, vi) mature equine oocytes, vii) capacitating and viii) non-capacitating medium. None of these triggered sperm release or hyperactivated motility. Interestingly, native FF was detrimental to sperm viability, an effect that was negated by heat inactivation, charcoal treatment and 30 kDa filtration alone or in combination. Moreover, sperm suspensions exposed to treated FF at pH 7.9 but not pH 7.4 showed Ca(2+)-dependent hypermotility. Fluo-4 AM staining of sperm showed elevated cytoplasmic Ca(2+) in hyperactivated stallion spermatozoa exposed to treated FF at pH 7.9 compared to a modest response in defined capacitating conditions at pH 7.9 and no response in treated FF at pH 7.4. Moreover, 1 h incubation in alkaline, treated FF induced protein tyrosine phosphorylation in 20% of spermatozoa. None of the conditions tested induced widespread release of sperm pre-bound to oviduct epithelium. However, the hyperactivating conditions did induce release of 70-120 spermatozoa per oviduct explant, of which 48% showed protein tyrosine phosphorylation and all were acrosome-intact, but capable of acrosomal exocytosis in response to calcium ionophore. We conclude that, in the presence of elevated pH and extracellular Ca(2+), a heat-resistant, hydrophilic, <30 kDa component of FF can trigger protein tyrosine phosphorylation, elevated cytoplasmic Ca(2+) and hyperactivated motility in stallion sperm, but infrequent release of sperm pre-bound to oviduct epithelium.
© 2015 Society for Reproduction and Fertility.
Publication Date: 2015-08-06 PubMed ID: 26242588DOI: 10.1530/REP-15-0178Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research explores the impact of different fluid conditions on the release and viability of horse spermatozoa (sperm cells) as they prepare for fertilization in the oviduct (a tube connecting the ovary to the uterus).
Understanding the Objective of the Research
The main objective of this research was to understand what factors can stimulate the release of sperm cells that are bound to the oviduct in preparation for fertilization. The researchers are particularly interested in the effects of fluid from different stages of the ovulatory cycle, as well as the physiological conditions such as pH level and calcium concentration.
Research Methodologies and Procedures
The research utilized a range of procedures to attain their objectives:
- Exposing horse sperm cells to various substances, including pre-ovulatory and post-ovulatory oviductal fluid, follicular fluid (FF), cumulus cells, mature equine oocytes, capacitating and non-capacitating medium.
- Investigating the viability and motility of the sperm cells under these different conditions.
- Assessing the effect of these fluids on sperm cells when treated with heat inactivation, charcoal treatment, and 30 kDa filtration.
- Evaluating the hyper motility achieved in different pH conditions, specifically pH 7.9 and 7.4.
Key Findings of the Study
The key findings of this research can be broken down as follows:
- None of the conditions tested triggered release or increased motility of the sperm cells.
- Untreated follicular fluid was found to be detrimental to sperm viability.
- The negative effect of follicular fluid on sperm viability could be negated via heat inactivating, charcoal treating, or filtration of certain molecule sizes.
- Sperm cells exposed to treated follicular fluid at pH 7.9 displayed calcium-dependent hypermotility, indicating that pH and calcium levels had an impact on sperm motility.
- There was no significant release of sperm cells pre-bound to oviduct epithelium under any of the conditions tested, but there was a minor release under hyperactivating conditions.
Conclusions from the Research
The research concludes that a constituent of follicular fluid, when heat-resistant and less than 30kDa, can trigger protein tyrosine phosphorylation, calcium elevation, and hyperactive motility in horse sperm cells. This reaction only occurs in alkaline conditions with elevated calcium levels. However, this stimulation does not lead to a significant release of sperm pre-bound to the oviduct epithelium.
Cite This Article
APA
Leemans B, Gadella BM, Stout TA, Nelis H, Hoogewijs M, Van Soom A.
(2015).
An alkaline follicular fluid fraction induces capacitation and limited release of oviduct epithelium-bound stallion sperm.
Reproduction, 150(3), 193-208.
https://doi.org/10.1530/REP-15-0178 Publication
Researcher Affiliations
- Department of ReproductionObstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, BelgiumDepartments of Farm Animal HealthBiochemistry and Cell BiologyEquine SciencesFaculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands baleeman.leemans@ugent.be.
- Department of ReproductionObstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, BelgiumDepartments of Farm Animal HealthBiochemistry and Cell BiologyEquine SciencesFaculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands Department of ReproductionObstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, BelgiumDepartments of Farm Animal HealthBiochemistry and Cell BiologyEquine SciencesFaculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
- Department of ReproductionObstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, BelgiumDepartments of Farm Animal HealthBiochemistry and Cell BiologyEquine SciencesFaculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands Department of ReproductionObstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, BelgiumDepartments of Farm Animal HealthBiochemistry and Cell BiologyEquine SciencesFaculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
- Department of ReproductionObstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, BelgiumDepartments of Farm Animal HealthBiochemistry and Cell BiologyEquine SciencesFaculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
- Department of ReproductionObstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, BelgiumDepartments of Farm Animal HealthBiochemistry and Cell BiologyEquine SciencesFaculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
- Department of ReproductionObstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, BelgiumDepartments of Farm Animal HealthBiochemistry and Cell BiologyEquine SciencesFaculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
MeSH Terms
- Acrosome Reaction
- Animals
- Calcium / metabolism
- Calcium Ionophores / pharmacology
- Cell Adhesion
- Epithelial Cells / metabolism
- Female
- Follicular Fluid / metabolism
- Horses / physiology
- Hydrogen-Ion Concentration
- Male
- Oviducts / metabolism
- Phosphorylation
- Sperm Capacitation / drug effects
- Sperm Motility / drug effects
- Spermatozoa / drug effects
- Spermatozoa / metabolism
- Time Factors
- Tissue Culture Techniques
- Tyrosine / metabolism
Citations
This article has been cited 5 times.- Lawson EF, Grupen CG, Baker MA, Aitken RJ, Swegen A, Pollard CL, Gibb Z. Conception and early pregnancy in the mare: lipidomics the unexplored frontier.. Reprod Fertil 2022 Jan 1;3(1):R1-R18.
- 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.
- Ruiz-Díaz S, Oseguera-López I, De La Cuesta-Díaz D, García-López B, Serres C, Sanchez-Calabuig MJ, Gutiérrez-Adán A, Perez-Cerezales S. The Presence of D-Penicillamine during the In Vitro Capacitation of Stallion Spermatozoa Prolongs Hyperactive-Like Motility and Allows for Sperm Selection by Thermotaxis.. Animals (Basel) 2020 Aug 21;10(9).
- 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.
- Kerns K, Zigo M, Drobnis EZ, Sutovsky M, Sutovsky P. Zinc ion flux during mammalian sperm capacitation.. Nat Commun 2018 May 25;9(1):2061.
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