Effects of glutamine, proline, histidine and betaine on post-thaw motility of stallion spermatozoa.
Abstract: The supplementation of the freezing diluent with 3 amino acids (glutamine, proline and histidine) and 1 amino acid-related compound (betaine) in preserving stallion spermatozoa diluted in INRA82 extender containing 2.5% (v/v) glycerol and 2% (v/v) egg yolk (control extender) during freezing and thawing was studied at 0, 40, 80, 120 and 160 mM in 20 split ejaculates (10 stallions x 2 ejaculates; Experiment 1). Glutamine and proline were studied at 0, 10, 20, 30, 40, 50, 60, 70 and 80 mM in 20 split ejaculates (10 stallions x 2 ejaculates; Experiment 2). In each experiment, spermatozoa were evaluated after thawing by computer automated sperm analyzer. The percentage of motile spermatozoa (faster than 30 microns/sec) was assessed. In addition, the velocity of the average path (VAP), the straight line velocity (VSL), the curvilinear velocity (VCL) and the amplitude of the lateral head displacement (ALH) were also measured. In Experiment 1, only glutamine (40 mM) significantly improved sperm motility (56.0% +/- 3.0 vs 49.7% +/- 1.6; P < 0.05) compared with the control extender, while velocities were unaffected at concentrations of 40 to 120 mM. However, at 160 mM, a significant decrease in motility and velocity was observed for all amino acids. In Experiment 2, motility in glutamine (range 41.1% +/- 3.8%; 42.4% +/- 3.6) and proline (43.0% +/- 3.7; 45.6% +/- 3.8) extenders compared with the control (34.7% +/- 1.6) was improved significantly (P < 0.05). Sperm velocity was improved at concentrations higher than 40 mM glutamine and 50 mM proline.
Publication Date: 2000-03-29 PubMed ID: 10734416DOI: 10.1016/s0093-691x(99)00120-xGoogle Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research study investigates the impact of amino acid supplementation on the motility of stallion spermatozoa after a process of freezing and thawing. The study found that among the amino acids tested, glutamine was most effective in enhancing sperm motility.
Objective of the Research
- The main objective of this research was to study the effects of supplementing the freezing diluent with three different types of amino acids (glutamine, proline, and histidine) and one amino acid-related compound (betaine) on the post-thaw motility of stallion spermatozoa.
Research Methodology
- The study was split into two experiments. For both experiments, 20 split ejaculates (10 stallions x 2 ejaculates each) were utilized.
- The amino acids were tested at different concentrations and evaluated after the thawing process, using a computer automated sperm analyzer.
- The measures of evaluation included the percentage of motile spermatozoa, and the velocity measures of the average path (VAP), the straight line velocity (VSL), the curvilinear velocity (VCL), and the amplitude of the lateral head displacement (ALH).
Findings of the Research
- In the first experiment, it was found that 40 mM of glutamine significantly enhanced sperm motility, but the velocities were not affected in concentrations ranging from 40 to 120 mM. However, at 160 mM concentration, the motility and velocity decreased significantly for all amino acids.
- In the second experiment, sperm motility was noticeably improved in glutamine and proline extenders compared to the control extender. Sperm velocity was improved at concentrations higher than 40 mM glutamine and 50 mM proline.
- Importantly, the study suggests that the amino acid glutamine, provided at the correct concentration, could be utilized as a supplement in the freezing process to improve post-thaw sperm motility.
Conclusion of the Research
- This study concluded that the amino acid glutamine could provide a significant improvement in the post-thaw motility of stallion spermatozoa. Further research is suggested to validate these findings and explore other amino acids that could potentially enhance post-thaw sperm motility.
Cite This Article
APA
Trimeche A, Yvon JM, Vidament M, Palmer E, Magistrini M.
(2000).
Effects of glutamine, proline, histidine and betaine on post-thaw motility of stallion spermatozoa.
Theriogenology, 52(1), 181-191.
https://doi.org/10.1016/s0093-691x(99)00120-x Publication
Researcher Affiliations
- INRA-Haras Nationaux, Unité PRMD, Nouzilly, France.
MeSH Terms
- Animals
- Betaine / pharmacology
- Cryopreservation
- Cryoprotective Agents
- Egg Yolk
- Glutamine / pharmacology
- Glycerol
- Histidine / pharmacology
- Horses
- Male
- Proline / pharmacology
- Semen Preservation
- Sperm Motility / drug effects
Citations
This article has been cited 13 times.- Balogun KB, Nicholls G, Sokunbi OA, Stewart KR. Cryoprotectant effects of natural honey on spermatozoa quality of pre-freezing and frozen-thawed boar semen. J Anim Sci 2023 Jan 3;101.
- Sandfoss MR, Cantrell J, Roberts BM, Reichling S. Cryopreservation of Sperm from an Endangered Snake with Tests of Post-Thaw Incubation in Caffeine. Animals (Basel) 2022 Jul 17;12(14).
- Santiago-Moreno J, Blesbois E. Functional Aspects of Seminal Plasma in Bird Reproduction. Int J Mol Sci 2020 Aug 7;21(16).
- Lugar DW, Krom WA, Mings PD, Stewart KR. Effects of supplemental betaine to semen extenders on semen quality in boars. Transl Anim Sci 2018 Apr;2(2):195-204.
- Longobardi V, Kosior MA, Pagano N, Fatone G, Staropoli A, Vassetti A, Vinale F, Campanile G, Gasparrini B. Changes in Bull Semen Metabolome in Relation to Cryopreservation and Fertility. Animals (Basel) 2020 Jun 19;10(6).
- Ugur MR, Dinh T, Hitit M, Kaya A, Topper E, Didion B, Memili E. Amino Acids of Seminal Plasma Associated With Freezability of Bull Sperm. Front Cell Dev Biol 2019;7:347.
- Santiago-Moreno J, Bernal B, Pérez-Cerezales S, Castaño C, Toledano-Díaz A, Esteso MC, Gutiérrez-Adán A, López-Sebastián A, Gil MG, Woelders H, Blesbois E. Seminal plasma amino acid profile in different breeds of chicken: Role of seminal plasma on sperm cryoresistance. PLoS One 2019;14(1):e0209910.
- Li MW, Vallelunga JM, Kinchen KL, Rink KL, Zarrabi J, Shamamian AO, Lloyd KC. IVF recovery of mutant mouse lines using sperm cryopreserved with mtg in cryovials. Cryo Letters 2014 Mar-Apr;35(2):145-53.
- Buonaiuto G, Federiconi A, Vecchiato CG, Benini E, Mordenti AL. Betaine Dietary Supplementation: Healthy Aspects in Human and Animal Nutrition. Antioxidants (Basel) 2025 Jun 23;14(7).
- Nuntapaitoon M, Tummaruk P, Suwimonteerabutr J. Supplementation of glutamine in a short-term boar semen extender during 17°C holding time enhances post-thaw sperm quality for cryopreservation. Porcine Health Manag 2024 Nov 11;10(1):50.
- Li C, Liu C, Chen Y, Zhao Y, Tan M, He B. Protective Effects of Betaine on Boar Sperm Quality during Liquid Storage and Transport. Animals (Basel) 2024 Sep 19;14(18).
- Kameni SL, Dlamini NH, Feugang JM. Exploring the full potential of sperm function with nanotechnology tools. Anim Reprod 2024;21(3):e20240033.
- Zhang Y, Ding N, Cao J, Zhang J, Liu J, Zhang C, Jiang L. Proteomics and Metabolic Characteristics of Boar Seminal Plasma Extracellular Vesicles Reveal Biomarker Candidates Related to Sperm Motility. J Proteome Res 2024 Sep 6;23(9):3764-3779.
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