College of Veterinary Science, Assam Agricultural University, Guwahati, Assam, India.
ICAR-National Research Centre on Pig, Guwahati, Assam, India.
Reprod Domest Anim. 2024 Aug;59(8):e14679. doi: 10.1111/rda.14679.
In vitro capacitation allows for a greater understanding of the mechanisms underlying fertilization and the development of improved reproductive techniques for improving fertility rates in porcine. Tyrodes albumin lactate pyruvate (TALP) and modified Krebs Ringers Broth (m-KRB) are two medias that are commonly used in research experiments to induce capacitation in boar spermatozoa (Cañón-Beltrán et al., Theriogenology, 198, 2023 and 231; Oberlender et al., Archivos de Medicina Veterinaria, 44, 2012 and 201; Sahoo et al., International Journal of Biological Macromolecules, 241, 2023 and 124502). Moreover, understanding the morphological and functional changes in boar spermatozoa at different hours of capacitation periods might aid in the development of novel techniques for improving sperm quality and increasing the litter size. This study was carried out to investigate the effect of Tyrode albumin lactate pyruvate and modified Krebs Ringers Broth media on in vitro capacitation of HD-K75 boar spermatozoa at three different periods of incubation. A total of 24 ejaculate from four clinically healthy, 10-12 months aged HD-K75 boars, maintained at ICAR-All India Coordinated Research Project (AICRP) on pig were selected. Semen was collected by 'Simple fist' method using a portable dummy. The semen samples having 200 mL volume, 103 × 10 spermatozoa/ml concentration and 70% initial motility were selected and split into two parts and suspended in TALP and m-KRB media, respectively, and incubated for 5 h at 37°C. Seminal parameters viz. sperm viability, plasma membrane integrity and acrosomal integrity were estimated in the samples at 0, 3 and 5 h of incubation. This study revealed that there was significant variation between media in live acrosome-reacted (p < .05) and HOST-reacted (p < .01) spermatozoa, while between capacitation periods significant (p < .01) variation was observed in hyperactivated spermatozoa, live acrosome-reacted spermatozoa, HOST-reacted spermatozoa, FITC-labelled PSA, extracellular protein and sperm cholesterol. Non-significant variation was observed in total phospholipid. TALP showed overall better consequence on sperm viability, plasma membrane and acrosomal integrity of boar spermatozoa. From this study, it could be concluded that both TALP and m-KRB media were virtuous to induce capacitation in HD-K75 boar spermatozoa. TALP media, however, had a better effect on sperm viability, plasma membrane and acrosomal integrity of boar spermatozoa. Out of the three different periods, 3 h capacitation period resulted in significantly (p < .01) higher incidence of sperm viability, plasma membrane and acrosomal integrity in HD-K75 boar spermatozoa.
在体外获能允许我们更深入地了解受精的机制，并开发出改进的生殖技术，以提高猪的生育能力。Tyrodes 白蛋白乳酸盐丙酮酸盐（TALP）和改良 Krebs Ringers 肉汤（m-KRB）是两种常用于研究实验中诱导猪精子获能的培养基（Cañón-Beltrán 等人，《生殖学》，198，2023 年和 231 页；Oberlender 等人，《兽医档案》，44，2012 年和 201 页；Sahoo 等人，《国际生物大分子杂志》，241，2023 年和 124502）。此外，了解不同获能期猪精子的形态和功能变化，可能有助于开发提高精子质量和增加产仔数的新技术。本研究旨在探讨 Tyrode 白蛋白乳酸盐丙酮酸盐和改良 Krebs Ringers 肉汤培养基对 HD-K75 公猪精子在三个不同孵育期的体外获能的影响。从四个临床健康、10-12 月龄的 HD-K75 公猪的 24 个精液中选择了 24 个，这些公猪均由 ICAR-All India Coordinated Research Project（AICRP）维持。使用便携式假阴茎通过“简单握拳”法采集精液。选择体积为 200ml、精子浓度为 103×10 个/ml、初始活力为 70%的精液样本，并将其分成两部分，分别悬浮在 TALP 和 m-KRB 培养基中，在 37°C 下孵育 5 小时。在孵育 0、3 和 5 小时时，分别评估样本中的精子活力、质膜完整性和顶体完整性等精液参数。本研究表明，两种培养基在活顶体反应（p<.05）和 HOST 反应（p<.01）精子之间存在显著差异，而在不同的获能期，超激活精子、活顶体反应精子、HOST 反应精子、FITC 标记 PSA、细胞外蛋白和精子胆固醇之间存在显著差异（p<.01）。总磷脂没有观察到显著变化。TALP 对猪精子的活力、质膜和顶体完整性总体表现出更好的效果。从这项研究中可以得出结论，TALP 和 m-KRB 培养基都能有效地诱导 HD-K75 公猪精子获能。然而，TALP 培养基对猪精子的活力、质膜和顶体完整性有更好的影响。在三个不同的时期中，3 小时的获能期可显著（p<.01）提高 HD-K75 公猪精子的活力、质膜和顶体完整性。
