Animal Biosciences and Biotechnology Laboratory, Beltsville Agricultural Research Center, USDA, Beltsville, MD 20705, USA.
Poult Sci. 2012 Aug;91(8):1990-6. doi: 10.3382/ps.2011-02028.
The ability to store turkey semen for 24 h in vitro without a significant loss in fertility would benefit the commercial turkey industry. We investigated the use of exogenous phosphatidylcholine to improve the viability, mobility, hydrolyzing ability, and fertility of stored turkey sperm. For experiment 1, semen was diluted 1:1 with extender containing 0.0 (control), 0.5, 2.5, or 10.0 mg/mL of phosphatidylcholine labeled with a fluorochrome and maintained under aerobic conditions for 24 h at 4°C. Semen aliquots were removed at 30-min intervals during the first 4 h and at 1-h intervals from 8 to 24 h of storage for fluorometric evaluation by flow cytometry. Turkey sperm incorporated labeled phosphatidylcholine in a dose-dependent manner during the first 12 h of storage (P<0.05). At 24 h of storage, phosphatidylcholine uptake increased 7.8-fold for the 0.5-mg treatment, 9.2-fold for the 2.5-mg treatment, and 6.7-fold for the 10-mg labeled phosphatidylcholine treatment. For experiment 2, turkey semen was diluted and stored as for experiment 1 except the phosphatidylcholine was unlabeled. After 24 h, the viability, mobility, hydrolyzing ability, and fertility of turkey sperm was assessed. Supplemental phosphatidylcholine did not improve (P>0.05) the viability or mobility of stored sperm. At 2.5 mg/mL, phosphatidylcholine improved the hydrolyzing ability of stored sperm compared with control or other phosphatidylcholine treatments (P<0.05). The mean fertility rate of eggs from hens inseminated with control semen was 33.5%±4.5. Use of 0.5, 2.5, or 10.0 mg phosphatidylcholine/mL improved (P<0.05) the fertility rates during the first 11 wk of egg production; higher fertility rates occurred with 2.5 mg phosphatidylcholine/mL compared with other phosphatidylcholine treatments for 5 of those 11 wk (P<0.05). We conclude that supplemental phosphatidylcholine appears to counteract the damaging effects of lipid peroxidation or enzymatic degradation during in vitro storage by providing exogenous phospholipids for incorporation into the turkey sperm plasma membrane.
将火鸡精液在体外储存 24 小时而不显著降低生育力的能力将有益于商业火鸡产业。我们研究了使用外源性磷脂酰胆碱来提高储存的火鸡精子的活力、运动性、水解能力和生育力。在实验 1 中,精液用含有 0.0(对照)、0.5、2.5 或 10.0 mg/mL 荧光标记磷脂酰胆碱的稀释液以 1:1 的比例稀释,并在有氧条件下于 4°C 下储存 24 小时。在储存的前 4 小时内每 30 分钟取一次精液样本,从 8 小时到 24 小时每 1 小时取一次样本,通过流式细胞术进行荧光评估。火鸡精子在储存的前 12 小时内以剂量依赖的方式摄取标记的磷脂酰胆碱(P<0.05)。在储存 24 小时时,0.5mg 处理组的磷脂酰胆碱摄取增加了 7.8 倍,2.5mg 处理组增加了 9.2 倍,10mg 标记磷脂酰胆碱处理组增加了 6.7 倍。在实验 2 中,除了磷脂酰胆碱未标记外,火鸡精液的稀释和储存与实验 1 相同。储存 24 小时后,评估火鸡精子的活力、运动性、水解能力和生育力。补充磷脂酰胆碱并没有提高(P>0.05)储存精子的活力或运动性。在 2.5mg/mL 时,与对照或其他磷脂酰胆碱处理相比,磷脂酰胆碱提高了储存精子的水解能力(P<0.05)。用对照精液授精母鸡的蛋的平均受精率为 33.5%±4.5。在产卵的前 11 周内,使用 0.5、2.5 或 10.0mg 磷脂酰胆碱/mL 提高了受精率(P<0.05);与其他磷脂酰胆碱处理相比,在 11 周中的 5 周中,2.5mg 磷脂酰胆碱/mL 组的受精率更高(P<0.05)。我们的结论是,补充磷脂酰胆碱似乎通过提供外源性磷脂来与体外储存过程中的脂质过氧化或酶促降解作用相抗衡,从而将其掺入火鸡精子质膜中。