Lactate as the sole energy substrate induces spontaneous acrosome reaction in viable stallion spermatozoa.
Abstract: Equine spermatozoa appear to differ from spermatozoa of other species in using oxidative phosphorylation preferentially over glycolysis. However, there is little information regarding effects of different energy sources on measured parameters in equine spermatozoa. Objective: To determine the effect of three individual energy substrates, glucose, pyruvate, and lactate, on motion characteristics, membrane integrity, and acrosomal status of stallion spermatozoa. Methods: Freshly ejaculated stallion spermatozoa were incubated with combinations of glucose (5 mm), pyruvate (10 mm), and lactate (10 mm) for 0.5 to 4 h. Response to calcium ionophore A23187 (5 μm) was used to evaluate capacitation status. Motility was evaluated using computer-assisted sperm analysis, and plasma membrane and acrosomal integrity were evaluated by flow cytometry. Results: Incubation with lactate alone for 2 h increased acrosomal sensitivity to A23187. Notably, incubation with lactate alone for 4 h induced a significant spontaneous increase in acrosome-reacted, membrane-intact (viable) spermatozoa, to approximately 50% of the live population, whereas no increase was seen with incubation in glucose or pyruvate alone. This acrosomal effect was observed in spermatozoa incubated at physiological pH as well as under alkaline conditions (medium pH approximately 8.5). Sperm motility declined concomitantly with the increase in acrosome-reacted spermatozoa. Sperm motility was significantly higher in pyruvate-only medium than in glucose or lactate. The addition of pyruvate to lactate-containing medium increased sperm motility but reduced the proportion of live acrosome-reacted spermatozoa in a dose-dependent fashion. Conclusions: This is the first study to demonstrate that incubation with a specific energy substrate, lactate, is associated with spontaneous acrosome reaction in spermatozoa. The proportion of live, acrosome-reacted spermatozoa obtained is among the highest reported for equine spermatozoa. Conclusions: These findings highlight the delicate control of key sperm functions, and may serve as a basis to increase our understanding of stallion sperm physiology.
© 2023 The Authors. Andrology published by Wiley Periodicals LLC on behalf of American Society of Andrology and European Academy of Andrology.
Publication Date: 2023-06-10 PubMed ID: 37300872DOI: 10.1111/andr.13479Google Scholar: Lookup
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Summary
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This research highlights the distinctive effect of different energy sources, specifically lactate, on the characteristics of stallion spermatozoa including motion, membrane integrity and acrosomal status. The main findings reveal that lactate, when used alone as an energy source, triggers a spontaneous reaction in sperm’s acrosome, a phenomenon not observed with glucose or pyruvate.
Objectives and Methods
- The primary objective of the study was to ascertain the influence of three distinct energy substrates (glucose, pyruvate, and lactate) on the motion characteristics, membrane integrity, and the acrosomal status of stallion spermatozoa.
- Fresh stallion spermatozoa, post-ejaculation, were incubated with individual energy substrates and their combinations in varied time durations ranging from 0.5 to 4 hours.
- Responses to calcium ionophore A23187 were evaluated to determine the capacitation status of the sperm, while computer-assisted sperm analysis was harnessed for assessing motility. Flow cytometry was employed to evaluate plasma membrane and acrosomal integrity.
Key Results
- An increased acrosomal sensitivity to A23187 was noticed when the sperm were incubated with lactate only, for 2 hours.
- A significant spontaneous increase in acrosome-reacted, yet viable spermatozoa, was observed when incubated for 4 hours with lactate alone. However, such an increase was not noted when the spermatozoa were incubated with glucose or pyruvate alone.
- The process was effective under both, physiological pH conditions and alkaline conditions (medium pH approximately 8.5).
- Declination of sperm motility was seen, coinciding with the increase in acrosome-reacted spermatozoa.
- The spermatozoa demonstrated higher motility in the pyruvate-only medium than in glucose or lactate. But adding pyruvate to lactate medium augmented sperm motility while reducing the proportion of live acrosome-reacted spermatozoa.
Conclusions
- The research results stipulate a critical finding that exposure to the energy substrate, lactate, is associated with a spontaneous acrosome reaction in spermatozoa.
- The proportion of viable, acrosome-reacted spermatozoa obtained in these conditions is one of the highest reported for equine spermatozoa.
- The findings underscore the delicate equilibrium of key sperm functions and can serve in enhancing our comprehension of stallion sperm physiology.
Cite This Article
APA
Ramírez-Agámez L, Hernández-Avilés C, Ortíz I, Love CC, Varner DD, Hinrichs K.
(2023).
Lactate as the sole energy substrate induces spontaneous acrosome reaction in viable stallion spermatozoa.
Andrology.
https://doi.org/10.1111/andr.13479 Publication
Researcher Affiliations
- Department of Veterinary Physiology and Pharmacology, School of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, Texas, USA.
- Department of Large Animal Clinical Sciences, School of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, Texas, USA.
- Department of Large Animal Clinical Sciences, School of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, Texas, USA.
- Department of Veterinary Physiology and Pharmacology, School of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, Texas, USA.
- Department of Large Animal Clinical Sciences, School of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, Texas, USA.
- Department of Large Animal Clinical Sciences, School of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, Texas, USA.
- Department of Veterinary Physiology and Pharmacology, School of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, Texas, USA.
- Department of Large Animal Clinical Sciences, School of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, Texas, USA.
- Department of Clinical Studies - New Bolton Center, School of Veterinary Medicine, University of Pennsylvania, Kennett Square, Pennsylvania, USA.
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Citations
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