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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2025-06-01
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| author | Yuting Kong Mengqian He Jun Gao Jiehuan Xu Naisheng Lu Caifeng Wu Lingwei Sun Jianjun Dai |
| author_facet | Yuting Kong Mengqian He Jun Gao Jiehuan Xu Naisheng Lu Caifeng Wu Lingwei Sun Jianjun Dai |
| author_sort | Yuting Kong |
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| description | 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 (<i>p</i> < 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; <i>p</i> < 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 (<i>p</i> < 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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| institution | Kabale University |
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| language | English |
| publishDate | 2025-06-01 |
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| series | Animals |
| spelling | doaj-art-2545faa25ae74930b86ca0bfadf966472025-08-20T03:28:32ZengMDPI AGAnimals2076-26152025-06-011513188510.3390/ani15131885Comparative Analysis of Classic Semen Extenders for Frozen–Thawed Boar SemenYuting Kong0Mengqian He1Jun Gao2Jiehuan Xu3Naisheng Lu4Caifeng Wu5Lingwei Sun6Jianjun Dai7College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, ChinaShanghai Municipal Key Laboratory of Agri-Genetics and Breeding, Institute of Animal Science and Veterinary Medicine, Shanghai Academy of Agricultural Sciences, Shanghai 201106, ChinaShanghai Municipal Key Laboratory of Agri-Genetics and Breeding, Institute of Animal Science and Veterinary Medicine, Shanghai Academy of Agricultural Sciences, Shanghai 201106, ChinaShanghai Municipal Key Laboratory of Agri-Genetics and Breeding, Institute of Animal Science and Veterinary Medicine, Shanghai Academy of Agricultural Sciences, Shanghai 201106, ChinaShanghai Municipal Key Laboratory of Agri-Genetics and Breeding, Institute of Animal Science and Veterinary Medicine, Shanghai Academy of Agricultural Sciences, Shanghai 201106, ChinaShanghai Municipal Key Laboratory of Agri-Genetics and Breeding, Institute of Animal Science and Veterinary Medicine, Shanghai Academy of Agricultural Sciences, Shanghai 201106, ChinaShanghai Municipal Key Laboratory of Agri-Genetics and Breeding, Institute of Animal Science and Veterinary Medicine, Shanghai Academy of Agricultural Sciences, Shanghai 201106, ChinaShanghai Municipal Key Laboratory of Agri-Genetics and Breeding, Institute of Animal Science and Veterinary Medicine, Shanghai Academy of Agricultural Sciences, Shanghai 201106, ChinaThe 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 (<i>p</i> < 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; <i>p</i> < 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 (<i>p</i> < 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.https://www.mdpi.com/2076-2615/15/13/1885boar semencryopreservationsemen extenderenergy metabolismspermatozoa motility |
| spellingShingle | Yuting Kong Mengqian He Jun Gao Jiehuan Xu Naisheng Lu Caifeng Wu Lingwei Sun Jianjun Dai Comparative Analysis of Classic Semen Extenders for Frozen–Thawed Boar Semen Animals boar semen cryopreservation semen extender energy metabolism spermatozoa motility |
| title | Comparative Analysis of Classic Semen Extenders for Frozen–Thawed Boar Semen |
| title_full | Comparative Analysis of Classic Semen Extenders for Frozen–Thawed Boar Semen |
| title_fullStr | Comparative Analysis of Classic Semen Extenders for Frozen–Thawed Boar Semen |
| title_full_unstemmed | Comparative Analysis of Classic Semen Extenders for Frozen–Thawed Boar Semen |
| title_short | Comparative Analysis of Classic Semen Extenders for Frozen–Thawed Boar Semen |
| title_sort | comparative analysis of classic semen extenders for frozen thawed boar semen |
| topic | boar semen cryopreservation semen extender energy metabolism spermatozoa motility |
| url | https://www.mdpi.com/2076-2615/15/13/1885 |
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