Chelucci Sara, Pasciu Valeria, Succu Sara, Addis Daniela, Leoni Giovanni G, Manca Maria E, Naitana Salvatore, Berlinguer Fiammetta
Department of Veterinary Medicine, University of Sassari, Sassari, Italy.
Department of Veterinary Medicine, University of Sassari, Sassari, Italy; Centro di Competenza Biodiversità Animale, Sassari, Italy.
Theriogenology. 2015 Apr 1;83(6):1064-74. doi: 10.1016/j.theriogenology.2014.12.012. Epub 2014 Dec 13.
Soybean lecithin may represent a suitable alternative to egg yolk for semen cryopreservation in livestock species. However, additional studies are needed to elucidate its effects on spermatozoa functional properties. Semen collected from five Sarda bucks was cryopreserved in Tris-based extender and glycerol (4% v:v) with different supplementations. In a preliminary experiment, different soybean lecithin concentrations were tested (1%-6% wt/vol) and results in terms of viability, percentages of progressive motile and rapid spermatozoa, and DNA integrity after thawing showed that the most effective concentration was 1%. In the second experiment, semen was frozen in a Tris-based extender with no supplementation (EXT), with 1% lecithin (EXT LC), and 20% egg yolk (EXT EY). The effectiveness of these extenders was also compared with a commercial extender. The EXT EY led to the highest viability and motility parameters after freezing and thawing (P < 0.0001). No significant differences were observed in intracellular ATP concentrations. Additional molecular features revealed that sperm functionality was affected in EXT EY, as demonstrated by lower DNA and acrosome integrity (P < 0.05), and higher lipid peroxidation compared with spermatozoa cryopreserved in EXT LC (P < 0.0001). Results obtained in the heterologous in vitro fertilization test showed that EXT LC better preserved spermatozoa functionality, as demonstrated by the higher fertilization rates compared with the other media (66.2 ± 4.5% for EXT LC vs. 32.7 ± 4.5%, 38.7 ± 4.5%, 39.6 ± 5.2% for EXT, EXT EY, and commercial extender; P < 0.01). The present study demonstrated that lecithin can be considered as a suitable alternative to egg yolk in goat semen cryopreservation, because it ensures higher fertilization rates and a better protection from membrane damage by cold shock.
大豆卵磷脂可能是家畜精液冷冻保存中蛋黄的合适替代品。然而,需要更多研究来阐明其对精子功能特性的影响。从五只撒丁岛公羊采集的精液在含甘油(4% v:v)的Tris基础稀释液中进行冷冻保存,并添加不同物质。在初步实验中,测试了不同浓度的大豆卵磷脂(1%-6% wt/vol),解冻后的活力、进行性运动精子和快速精子百分比以及DNA完整性结果表明,最有效的浓度为1%。在第二个实验中,精液在无添加物的Tris基础稀释液(EXT)、含1%卵磷脂的Tris基础稀释液(EXT LC)和含20%蛋黄的Tris基础稀释液(EXT EY)中冷冻。还将这些稀释液的效果与一种商业稀释液进行了比较。冷冻和解冻后,EXT EY的活力和运动参数最高(P < 0.0001)。细胞内ATP浓度未观察到显著差异。其他分子特征表明,与在EXT LC中冷冻保存的精子相比,EXT EY中的精子功能受到影响,表现为DNA和顶体完整性较低(P < 0.05),脂质过氧化程度较高(P < 0.0001)。在异源体外受精试验中获得的结果表明,EXT LC能更好地保存精子功能,与其他培养基相比受精率更高(EXT LC为66.2 ± 4.5%,EXT、EXT EY和商业稀释液分别为32.7 ± 4.5%、38.7 ± 4.5%、39.6 ± 5.2%;P < 0.01)。本研究表明,卵磷脂可被视为山羊精液冷冻保存中蛋黄的合适替代品,因为它能确保更高的受精率,并更好地保护精子免受冷休克造成的膜损伤。