Capacitation-like changes in equine spermatozoa following cryopreservation.
Abstract: The primary objective of this study was to assess plasma membrane characteristics and activation of signal transduction pathways in equine spermatozoa during both in vitro capacitation and cryopreservation. Significant plasma membrane restructuring, as assessed by measurement of plasma membrane lipid disorder and phospholipid scrambling, was not observed until after cryopreservation and subsequent thawing (P < 0.05). Although in vitro capacitated cells also displayed increased plasma membrane lipid disorder and phospholipid scrambling (P < 0.05), it appeared that regulation of these events in in vitro capacitated versus cryopreserved equine spermatozoa was not identical. Addition of 5 microM staurosporine to the capacitation media reduced plasma membrane phospholipid scrambling (P < 0.05), but supplementation to the freezing extender prior to cryopreservation did not. Furthermore, progesterone was able to induce a greater degree of acrosomal exocytosis in in vitro capacitated versus frozen/thawed spermatozoa. Expression of phospholipid scramblase, a protein thought to be important in plasma membrane phospholipid scrambling, did not differ between treatments. Comparison of protein tyrosine phosphorylation patterns between in vitro capacitated and cryopreserved cells demonstrated a divergence in signal transduction. Cellular signaling in in vitro capacitated equine spermatozoa appeared to be in part dependent on activation of the cAMP/PKA pathway, whereas signaling in cryopreserved cells seemed to proceed predominantly through alternative pathways. Taken together, these data support the idea that capacitation and "cryocapacitation" are not equivalent processes.
Publication Date: 2005-10-12 PubMed ID: 16225914DOI: 10.1016/j.theriogenology.2005.08.022Google Scholar: Lookup
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- Journal Article
- Research Support
- N.I.H.
- Extramural
- Research Support
- Non-U.S. Gov't
- Research Support
- U.S. Gov't
- Non-P.H.S.
Summary
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The research studies the effects of capacitation and cryopreservation on equine sperm cells. It was found that both processes cause alterations to the plasma membrane of the sperm cells, implicating changes in signal transduction pathways, but these effects are regulated differently.
Objective and Methodology
- The aim of this research was to examine plasma membrane characteristics and the activation of the signal transduction pathways in equine sperm during both in vitro capacitation and cryopreservation. Capacitation is a process that sperm undergo to acquire the ability to fertilize an egg, whereas cryopreservation is the process of preserving biological material at very low temperatures.
- The researchers measured parameters such as plasma membrane lipid disorder and phospholipid scrambling to assess the restructuring of the plasma membrane.
Findings
- Significant restructuring of the plasma membrane was observed only after cryopreservation and thawing.
- Cryopreserved cells and in vitro capacitated cells both exhibited increased plasma membrane lipid disorder and phospholipid scrambling. However, the regulation of these events in both processes appeared to be distinct, indicating different mechanisms in play.
- Supplementing the capacitation media with staurosporine, a protein kinase inhibitor, reduced the plasma membrane phospholipid scrambling in capacitated cells but didn’t yield the same results during cryopreservation.
- Progesterone induced a greater degree of acrosomal exocytosis (the process by which the sperm releases enzymes to penetrate the egg) in capacitated cells compared to frozen/thawed sperm cells.
- The expression of phospholipid scramblase, a protein that contributes to plasma membrane phospholipid scrambling, remained unchanged across all treatments.
- In vitro capacitated and cryopreserved cells showed different protein tyrosine phosphorylation patterns, indicating a divergence in signal transduction.
- In vitro capacitated equine sperm relies partly on the activation of the cAMP/PKA pathway, while cryopreserved cells seem to utilize alternative pathways for signaling.
Conclusion
- The results suggest that capacitation and “cryocapacitation” are different processes.
- Understanding these differences could help refining the techniques used in assisted reproduction in horses and possibly enhance their success rate.
Cite This Article
APA
Thomas AD, Meyers SA, Ball BA.
(2005).
Capacitation-like changes in equine spermatozoa following cryopreservation.
Theriogenology, 65(8), 1531-1550.
https://doi.org/10.1016/j.theriogenology.2005.08.022 Publication
Researcher Affiliations
- Department of Population, Health, and Reproduction, School of Veterinary Medicine, University of California, One Shields Avenue, Davis, CA 95616, USA.
MeSH Terms
- Animals
- Cell Membrane / drug effects
- Cell Membrane / physiology
- Cryopreservation / methods
- Cryopreservation / veterinary
- Cryoprotective Agents / pharmacology
- Enzyme Inhibitors / pharmacology
- Horses
- In Vitro Techniques
- Male
- Phospholipid Transfer Proteins / metabolism
- Semen Preservation / adverse effects
- Semen Preservation / veterinary
- Signal Transduction
- Sperm Capacitation / physiology
- Spermatozoa / metabolism
- Staurosporine / pharmacology
Grant Funding
- C06 RR-12088-01 / NCRR NIH HHS
Citations
This article has been cited 4 times.- 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).
- Takeuchi H, Nishioka M, Maezawa T, Kitano Y, Terada-Yoshikawa K, Tachibana R, Kato M, Hyon SH, Gen Y, Tanaka K, Toriyabe K, Nii M, Kondo E, Ikeda T. Carboxylated Poly-l-Lysine as a Macromolecular Cryoprotective Agent Enables the Development of Defined and Xeno-Free Human Sperm Cryopreservation Reagents.. Cells 2021 Jun 8;10(6).
- Patel A, Saxena A, Swain DK, Yadav D, Yadav SS, Kumar A, Kumar A. Effect of supplementation of butylated hydroxytoluene on post-thaw sperm viability, motility and membrane integrity of Hariana bulls.. Vet World 2015 Jun;8(6):808-12.
- 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.
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