A defined medium supports changes consistent with capacitation in stallion sperm, as evidenced by increases in protein tyrosine phosphorylation and high rates of acrosomal exocytosis.
Abstract: Efficient in vitro capacitation of stallion sperm has not yet been achieved, as suggested by low sperm penetration rates reported in in vitro fertilization (IVF) studies. Our objectives were to evaluate defined incubation conditions that would support changes consistent with capacitation in stallion sperm. Protein tyrosine phosphorylation events and the ability of sperm to undergo acrosomal exocytosis under various incubation conditions were used as end points for capacitation. Sperm incubated 4-6h in modified Whitten's (MW) with the addition of 25 mM NaHCO3 and 7 mg/mL BSA (capacitating medium) yielded high rates of protein tyrosine phosphorylation. Either HCO3(-) or BSA was required to support these changes, with the combination of both providing the most intense results. When a membrane-permeable form of cAMP and a phosphodiesterase inhibitor (IBMX) were added to MW in the absence of HCO3(-) and BSA, the tyrosine phosphorylation results obtained in our capacitating conditions could not be replicated, suggesting either effects apart from cAMP were responsible for tyrosine phosphorylation, or that stallion sperm might respond differently to these reagents as compared to sperm from other mammals. Sperm incubation in capacitating conditions was also associated with high percentages (P<or=0.001) of acrosomal exocytosis upon exposure to progesterone (44.6%) or calcium ionophore (51.6%), as compared to sperm incubated in medium devoid of BSA and NaHCO3. Our results were novel in that we report protein tyrosine phosphorylation in stallion sperm incubated in defined conditions, coupled with significant percentages of acrosome reacted sperm. The continuation of these studies might help to elucidate the conditions and pathways supporting sperm capacitation in the horse.
Publication Date: 2008-02-01 PubMed ID: 18242679DOI: 10.1016/j.theriogenology.2007.11.016Google 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.
- Journal Article
- Research Support
- Non-U.S. Gov't
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.
The research investigates the effective in vitro capacitation of stallion sperm. Researchers studied the impact of specific incubation conditions, observing increases in protein tyrosine phosphorylation and higher rates of acrosomal exocytosis—indicators of successful sperm capacitation.
Objective and Methodology
- The primary goal was to explore defined incubation conditions to support sperm capacitation in stallions, an area where efficient in vitro capacitation remains a challenge.
- End points for capacitation were based on protein tyrosine phosphorylation events and the sperm’s capability to undergo acrosomal exocytosis.
- The researchers incubated sperm for 4-6 hours in modified Whitten’s (MW) medium supplemented with 25 mM NaHCO3 and 7 mg/mL BSA (capacitating medium).
Results
- High rates of protein tyrosine phosphorylation were observed in the sperm incubated under the capacitating conditions.
- The combination of both HCO3(-) and BSA provided the most intense results; without them, the phosphorylation results could not be replicated.
- The implementation of a membrane-permeable form of cAMP and a phosphodiesterase inhibitor (IBMX) could not mimic the effects of the capacitating conditions either.
- These findings imply that factors beyond cAMP contribute to tyrosine phosphorylation or stallion sperm reacts differently to these agents compared to other mammalian sperm.
Evaluation of Acrosomal Exocytosis
- Sperm incubation under capacitating conditions revealed high percentages of acrosomal exocytosis upon exposure to progesterone (44.6%) or calcium ionophore (51.6%).
- These results significantly outperformed those of sperm incubated in a medium without BSA and NaHCO3.
Significance
- This research is groundbreaking in reporting protein tyrosine phosphorylation and significant percentages of acrosome reacted sperm in stallions under defined incubation conditions.
- Continuation of these studies may assist in illuminating the conditions and pathways that support sperm capacitation in horses.
Cite This Article
APA
McPartlin LA, Littell J, Mark E, Nelson JL, Travis AJ, Bedford-Guaus SJ.
(2008).
A defined medium supports changes consistent with capacitation in stallion sperm, as evidenced by increases in protein tyrosine phosphorylation and high rates of acrosomal exocytosis.
Theriogenology, 69(5), 639-650.
https://doi.org/10.1016/j.theriogenology.2007.11.016 Publication
Researcher Affiliations
- Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, United States.
MeSH Terms
- 1-Methyl-3-isobutylxanthine / pharmacology
- Acrosome Reaction / drug effects
- Acrosome Reaction / physiology
- Animals
- Bicarbonates / pharmacology
- Bucladesine / pharmacology
- Calcimycin / pharmacology
- Culture Media
- Cyclic AMP / physiology
- Horses / metabolism
- Horses / physiology
- Immunoblotting / veterinary
- Ionophores / pharmacology
- Male
- Phosphodiesterase Inhibitors / pharmacology
- Phosphorylation / drug effects
- Progesterone / pharmacology
- Serum Albumin, Bovine / pharmacology
- Sperm Capacitation / drug effects
- Sperm Capacitation / physiology
- Spermatozoa / metabolism
- Spermatozoa / physiology
- Tyrosine / metabolism
Citations
This article has been cited 9 times.- Aparicio IM, Rojo-Domínguez P, Castillejo-Rufo A, Peña FJ, Tapia JA. The Autophagy Marker LC3 Is Processed during the Sperm Capacitation and the Acrosome Reaction and Translocates to the Acrosome Where It Colocalizes with the Acrosomal Membranes in Horse Spermatozoa.. Int J Mol Sci 2023 Jan 4;24(2).
- Orsolini MF, Meyers SA, Dini P. An Update on Semen Physiology, Technologies, and Selection Techniques for the Advancement of In Vitro Equine Embryo Production: Section I.. Animals (Basel) 2021 Nov 13;11(11).
- Denisenko V, Chistyakova I, Volkova N, Volkova L, Iolchiev B, Kuzmina T. The Modulation of Functional Status of Bovine Spermatozoa by Progesterone.. Animals (Basel) 2021 Jun 15;11(6).
- Gimeno BF, Bariani MV, Laiz-Quiroga L, Martínez-León E, Von-Meyeren M, Rey O, Mutto AÁ, Osycka-Salut CE. Effects of In Vitro Interactions of Oviduct Epithelial Cells with Frozen-Thawed Stallion Spermatozoa on Their Motility, Viability and Capacitation Status.. Animals (Basel) 2021 Jan 3;11(1).
- Bucci D, Spinaci M, Galeati G, Tamanini C. Different approaches for assessing sperm function.. Anim Reprod 2020 May 22;16(1):72-80.
- 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.
- Fayrer-Hosken R, Stanley A, Hill N, Heusner G, Christian M, De La Fuente R, Baumann C, Jones L. Effect of feeding fescue seed containing ergot alkaloid toxins on stallion spermatogenesis and sperm cells.. Reprod Domest Anim 2012 Dec;47(6):1017-26.
- Filannino A, Stout TA, Gadella BM, Sostaric E, Pizzi F, Colenbrander B, Dell'Aquila ME, Minervini F. Dose-response effects of estrogenic mycotoxins (zearalenone, alpha- and beta-zearalenol) on motility, hyperactivation and the acrosome reaction of stallion sperm.. Reprod Biol Endocrinol 2011 Oct 5;9:134.
- McPartlin LA, Visconti PE, Bedford-Guaus SJ. Guanine-nucleotide exchange factors (RAPGEF3/RAPGEF4) induce sperm membrane depolarization and acrosomal exocytosis in capacitated stallion sperm.. Biol Reprod 2011 Jul;85(1):179-88.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
