Comparative Analysis of Classic Semen Extenders for Frozen–Thawed Boar Semen
The physiological functions of mammalian sperm, such as motility, hyperactivation, and capacitation, require substantial energy. This study investigates the effects of two classic cryopreservation extenders—TCG (tris-citrate-glucose) and LEY (lactose-egg yolk)—on the energy metabolism of frozen–thaw...
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| Published in | Animals (Basel) Vol. 15; no. 13; p. 1885 |
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| Main Authors | , , , , , , , |
| Format | Journal Article |
| Language | English |
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26.06.2025
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| ISSN | 2076-2615 2076-2615 |
| DOI | 10.3390/ani15131885 |
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| Abstract | The physiological functions of mammalian sperm, such as motility, hyperactivation, and capacitation, require substantial energy. This study investigates the effects of two classic cryopreservation extenders—TCG (tris-citrate-glucose) and LEY (lactose-egg yolk)—on the energy metabolism of frozen–thawed boar semen. By comparing the quality indicators, key metabolite levels, and the activities of critical enzymes involved in glycolysis and the tricarboxylic acid cycle, we aim to understand how these different semen extenders influence the spermatozoa vitality of frozen–thawed boar semen. Following thawing, the LEY-cryopreserved sperm demonstrated significantly elevated motility parameters (viability, VCL, VSL, and VAP) and enhanced plasma membrane and acrosomal integrity compared with the TCG group (p < 0.05), though both cryopreserved groups exhibited significantly reduced performance relative to fresh semen controls. Cryopreservation markedly reduced intracellular adenosine triphosphate (ATP), pyruvate, and acetyl coenzyme A (A-CoA) levels (fresh > LEY > TCG; p < 0.05). The LEY-preserved spermatozoa retained higher activities of glycolysis-related enzymes (phosphofructokinase, PFK; pyruvate kinase, PK) compared with the TCG group, which, in turn, showed elevated lactate dehydrogenase (LDH) activity. Critically, TCG-suppressed pyruvate dehydrogenase (PDH) activity (p < 0.05) coincided with diminished A-CoA, indicating impaired mitochondrial oxidative phosphorylation. These results demonstrate LEY’s superior preservation of motility and membrane stability but highlight cryodamage-induced energy metabolism dysregulation, particularly TCG’s disruption of the glycolysis–TCA cycle coordination essential for spermatozoa function. In conclusion, the choice of semen extender has a significant impact on the energy metabolism and overall quality of frozen–thawed semen, highlighting the importance of optimizing cryopreservation protocols for improved spermatozoa viability and functionality. |
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| AbstractList | The choice of semen extender is crucial for the in vitro preservation of mammalian sperm. This study compares two classic semen extenders for frozen–thawed boar semen—TCG (tris-citrate-glucose) and LEY (lactose-egg yolk)—to evaluate their effects on spermatozoa motility, structural integrity, and energy metabolism after thawing. The results indicate that the LEY-based semen extender significantly outperforms the TCG-based extender in maintaining the quality indicators of thawed semen, providing important insights for optimizing the cryopreservation techniques of boar semen. The choice of semen extender is crucial for the in vitro preservation of mammalian sperm. This study compares two classic semen extenders for frozen–thawed boar semen—TCG (tris-citrate-glucose) and LEY (lactose-egg yolk)—to evaluate their effects on spermatozoa motility, structural integrity, and energy metabolism after thawing. The results indicate that the LEY-based semen extender significantly outperforms the TCG-based extender in maintaining the quality indicators of thawed semen, providing important insights for optimizing the cryopreservation techniques of boar semen. The physiological functions of mammalian sperm, such as motility, hyperactivation, and capacitation, require substantial energy. This study investigates the effects of two classic cryopreservation extenders—TCG (tris-citrate-glucose) and LEY (lactose-egg yolk)—on the energy metabolism of frozen–thawed boar semen. By comparing the quality indicators, key metabolite levels, and the activities of critical enzymes involved in glycolysis and the tricarboxylic acid cycle, we aim to understand how these different semen extenders influence the spermatozoa vitality of frozen–thawed boar semen. Following thawing, the LEY-cryopreserved sperm demonstrated significantly elevated motility parameters (viability, VCL, VSL, and VAP) and enhanced plasma membrane and acrosomal integrity compared with the TCG group ( p < 0.05), though both cryopreserved groups exhibited significantly reduced performance relative to fresh semen controls. Cryopreservation markedly reduced intracellular adenosine triphosphate (ATP), pyruvate, and acetyl coenzyme A (A-CoA) levels (fresh > LEY > TCG; p < 0.05). The LEY-preserved spermatozoa retained higher activities of glycolysis-related enzymes (phosphofructokinase, PFK; pyruvate kinase, PK) compared with the TCG group, which, in turn, showed elevated lactate dehydrogenase (LDH) activity. Critically, TCG-suppressed pyruvate dehydrogenase (PDH) activity ( p < 0.05) coincided with diminished A-CoA, indicating impaired mitochondrial oxidative phosphorylation. These results demonstrate LEY’s superior preservation of motility and membrane stability but highlight cryodamage-induced energy metabolism dysregulation, particularly TCG’s disruption of the glycolysis–TCA cycle coordination essential for spermatozoa function. In conclusion, the choice of semen extender has a significant impact on the energy metabolism and overall quality of frozen–thawed semen, highlighting the importance of optimizing cryopreservation protocols for improved spermatozoa viability and functionality. The physiological functions of mammalian sperm, such as motility, hyperactivation, and capacitation, require substantial energy. This study investigates the effects of two classic cryopreservation extenders—TCG (tris-citrate-glucose) and LEY (lactose-egg yolk)—on the energy metabolism of frozen–thawed boar semen. By comparing the quality indicators, key metabolite levels, and the activities of critical enzymes involved in glycolysis and the tricarboxylic acid cycle, we aim to understand how these different semen extenders influence the spermatozoa vitality of frozen–thawed boar semen. Following thawing, the LEY-cryopreserved sperm demonstrated significantly elevated motility parameters (viability, VCL, VSL, and VAP) and enhanced plasma membrane and acrosomal integrity compared with the TCG group (p < 0.05), though both cryopreserved groups exhibited significantly reduced performance relative to fresh semen controls. Cryopreservation markedly reduced intracellular adenosine triphosphate (ATP), pyruvate, and acetyl coenzyme A (A-CoA) levels (fresh > LEY > TCG; p < 0.05). The LEY-preserved spermatozoa retained higher activities of glycolysis-related enzymes (phosphofructokinase, PFK; pyruvate kinase, PK) compared with the TCG group, which, in turn, showed elevated lactate dehydrogenase (LDH) activity. Critically, TCG-suppressed pyruvate dehydrogenase (PDH) activity (p < 0.05) coincided with diminished A-CoA, indicating impaired mitochondrial oxidative phosphorylation. These results demonstrate LEY’s superior preservation of motility and membrane stability but highlight cryodamage-induced energy metabolism dysregulation, particularly TCG’s disruption of the glycolysis–TCA cycle coordination essential for spermatozoa function. In conclusion, the choice of semen extender has a significant impact on the energy metabolism and overall quality of frozen–thawed semen, highlighting the importance of optimizing cryopreservation protocols for improved spermatozoa viability and functionality. The physiological functions of mammalian sperm, such as motility, hyperactivation, and capacitation, require substantial energy. This study investigates the effects of two classic cryopreservation extenders-TCG (tris-citrate-glucose) and LEY (lactose-egg yolk)-on the energy metabolism of frozen-thawed boar semen. By comparing the quality indicators, key metabolite levels, and the activities of critical enzymes involved in glycolysis and the tricarboxylic acid cycle, we aim to understand how these different semen extenders influence the spermatozoa vitality of frozen-thawed boar semen. Following thawing, the LEY-cryopreserved sperm demonstrated significantly elevated motility parameters (viability, VCL, VSL, and VAP) and enhanced plasma membrane and acrosomal integrity compared with the TCG group ( < 0.05), though both cryopreserved groups exhibited significantly reduced performance relative to fresh semen controls. Cryopreservation markedly reduced intracellular adenosine triphosphate (ATP), pyruvate, and acetyl coenzyme A (A-CoA) levels (fresh > LEY > TCG; < 0.05). The LEY-preserved spermatozoa retained higher activities of glycolysis-related enzymes (phosphofructokinase, PFK; pyruvate kinase, PK) compared with the TCG group, which, in turn, showed elevated lactate dehydrogenase (LDH) activity. Critically, TCG-suppressed pyruvate dehydrogenase (PDH) activity ( < 0.05) coincided with diminished A-CoA, indicating impaired mitochondrial oxidative phosphorylation. These results demonstrate LEY's superior preservation of motility and membrane stability but highlight cryodamage-induced energy metabolism dysregulation, particularly TCG's disruption of the glycolysis-TCA cycle coordination essential for spermatozoa function. In conclusion, the choice of semen extender has a significant impact on the energy metabolism and overall quality of frozen-thawed semen, highlighting the importance of optimizing cryopreservation protocols for improved spermatozoa viability and functionality. The physiological functions of mammalian sperm, such as motility, hyperactivation, and capacitation, require substantial energy. This study investigates the effects of two classic cryopreservation extenders-TCG (tris-citrate-glucose) and LEY (lactose-egg yolk)-on the energy metabolism of frozen-thawed boar semen. By comparing the quality indicators, key metabolite levels, and the activities of critical enzymes involved in glycolysis and the tricarboxylic acid cycle, we aim to understand how these different semen extenders influence the spermatozoa vitality of frozen-thawed boar semen. Following thawing, the LEY-cryopreserved sperm demonstrated significantly elevated motility parameters (viability, VCL, VSL, and VAP) and enhanced plasma membrane and acrosomal integrity compared with the TCG group (p < 0.05), though both cryopreserved groups exhibited significantly reduced performance relative to fresh semen controls. Cryopreservation markedly reduced intracellular adenosine triphosphate (ATP), pyruvate, and acetyl coenzyme A (A-CoA) levels (fresh > LEY > TCG; p < 0.05). The LEY-preserved spermatozoa retained higher activities of glycolysis-related enzymes (phosphofructokinase, PFK; pyruvate kinase, PK) compared with the TCG group, which, in turn, showed elevated lactate dehydrogenase (LDH) activity. Critically, TCG-suppressed pyruvate dehydrogenase (PDH) activity (p < 0.05) coincided with diminished A-CoA, indicating impaired mitochondrial oxidative phosphorylation. These results demonstrate LEY's superior preservation of motility and membrane stability but highlight cryodamage-induced energy metabolism dysregulation, particularly TCG's disruption of the glycolysis-TCA cycle coordination essential for spermatozoa function. In conclusion, the choice of semen extender has a significant impact on the energy metabolism and overall quality of frozen-thawed semen, highlighting the importance of optimizing cryopreservation protocols for improved spermatozoa viability and functionality.The physiological functions of mammalian sperm, such as motility, hyperactivation, and capacitation, require substantial energy. This study investigates the effects of two classic cryopreservation extenders-TCG (tris-citrate-glucose) and LEY (lactose-egg yolk)-on the energy metabolism of frozen-thawed boar semen. By comparing the quality indicators, key metabolite levels, and the activities of critical enzymes involved in glycolysis and the tricarboxylic acid cycle, we aim to understand how these different semen extenders influence the spermatozoa vitality of frozen-thawed boar semen. Following thawing, the LEY-cryopreserved sperm demonstrated significantly elevated motility parameters (viability, VCL, VSL, and VAP) and enhanced plasma membrane and acrosomal integrity compared with the TCG group (p < 0.05), though both cryopreserved groups exhibited significantly reduced performance relative to fresh semen controls. Cryopreservation markedly reduced intracellular adenosine triphosphate (ATP), pyruvate, and acetyl coenzyme A (A-CoA) levels (fresh > LEY > TCG; p < 0.05). The LEY-preserved spermatozoa retained higher activities of glycolysis-related enzymes (phosphofructokinase, PFK; pyruvate kinase, PK) compared with the TCG group, which, in turn, showed elevated lactate dehydrogenase (LDH) activity. Critically, TCG-suppressed pyruvate dehydrogenase (PDH) activity (p < 0.05) coincided with diminished A-CoA, indicating impaired mitochondrial oxidative phosphorylation. These results demonstrate LEY's superior preservation of motility and membrane stability but highlight cryodamage-induced energy metabolism dysregulation, particularly TCG's disruption of the glycolysis-TCA cycle coordination essential for spermatozoa function. In conclusion, the choice of semen extender has a significant impact on the energy metabolism and overall quality of frozen-thawed semen, highlighting the importance of optimizing cryopreservation protocols for improved spermatozoa viability and functionality. |
| Audience | Academic |
| Author | Wu, Caifeng Dai, Jianjun He, Mengqian Kong, Yuting Lu, Naisheng Xu, Jiehuan Gao, Jun Sun, Lingwei |
| AuthorAffiliation | 4 Shanghai Engineering Research Center of Breeding Pig, Shanghai 201302, China 3 Key Laboratory of Livestock and Poultry Resources (Pig) Evaluation and Utilization, Ministry of Agriculture and Rural Affairs, Shanghai 201106, China 2 Shanghai Municipal Key Laboratory of Agri-Genetics and Breeding, Institute of Animal Science and Veterinary Medicine, Shanghai Academy of Agricultural Sciences, Shanghai 201106, China; he1037247863@163.com (M.H.); gaojun@saas.sh.cn (J.G.); jiehuanxu810@163.com (J.X.); lunaisheng@saas.sh.cn (N.L.); wucaifengwcf@163.com (C.W.) 1 College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China; kytdyx2022@163.com |
| AuthorAffiliation_xml | – name: 4 Shanghai Engineering Research Center of Breeding Pig, Shanghai 201302, China – name: 2 Shanghai Municipal Key Laboratory of Agri-Genetics and Breeding, Institute of Animal Science and Veterinary Medicine, Shanghai Academy of Agricultural Sciences, Shanghai 201106, China; he1037247863@163.com (M.H.); gaojun@saas.sh.cn (J.G.); jiehuanxu810@163.com (J.X.); lunaisheng@saas.sh.cn (N.L.); wucaifengwcf@163.com (C.W.) – name: 3 Key Laboratory of Livestock and Poultry Resources (Pig) Evaluation and Utilization, Ministry of Agriculture and Rural Affairs, Shanghai 201106, China – name: 1 College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China; kytdyx2022@163.com |
| Author_xml | – sequence: 1 givenname: Yuting surname: Kong fullname: Kong, Yuting – sequence: 2 givenname: Mengqian orcidid: 0009-0000-6241-8291 surname: He fullname: He, Mengqian – sequence: 3 givenname: Jun orcidid: 0000-0002-2372-4445 surname: Gao fullname: Gao, Jun – sequence: 4 givenname: Jiehuan surname: Xu fullname: Xu, Jiehuan – sequence: 5 givenname: Naisheng surname: Lu fullname: Lu, Naisheng – sequence: 6 givenname: Caifeng surname: Wu fullname: Wu, Caifeng – sequence: 7 givenname: Lingwei orcidid: 0000-0002-3438-6584 surname: Sun fullname: Sun, Lingwei – sequence: 8 givenname: Jianjun surname: Dai fullname: Dai, Jianjun |
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| Keywords | boar semen spermatozoa motility cryopreservation energy metabolism semen extender |
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| Snippet | The physiological functions of mammalian sperm, such as motility, hyperactivation, and capacitation, require substantial energy. This study investigates the... The choice of semen extender is crucial for the in vitro preservation of mammalian sperm. This study compares two classic semen extenders for frozen–thawed... |
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| SubjectTerms | Analysis boar semen Cryopreservation Energy energy metabolism Enzymes Ethical aspects Glucose Kinases Lactose Metabolism Metabolites Motility Penicillin Physiological aspects Quality control Reagents semen extender spermatozoa motility |
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| Title | Comparative Analysis of Classic Semen Extenders for Frozen–Thawed Boar Semen |
| URI | https://www.ncbi.nlm.nih.gov/pubmed/40646783 https://www.proquest.com/docview/3229134155 https://www.proquest.com/docview/3229626746 https://pubmed.ncbi.nlm.nih.gov/PMC12248550 https://doaj.org/article/2545faa25ae74930b86ca0bfadf96647 |
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