摘要
The quality of the separated fractions in sex-sorted semen is very important for the success of the artificial insemination. This study aimed to evaluate some in vitro characteristics (DNA quantity, kinematic parameters and enzymes activity) of X- and Y-bearing ram spermatozoa sorted by bovine serum albumin (BSA) column and toll-like receptors (TLR)7/8 ligand R848. The ejaculates from six rams were collected by artificial vagina and subjected to a computer-assisted semen analysis (CASA). Total motility and percentage of the sperms with rapid and medium progressivity or non-progressivity in whole ejaculates and in X and Y fractions were analyzed. Activity of the enzymes ALP, GGT, CK, LDH and accumulation of lactate in the seminal plasma of ejaculates and in the environmental fluid of sexed spermatozoa were measured by biochemical analyzer. DNA was isolated from precipitated spermatozoa, and its quantity was measured. For both protocols the DNA mass from X-bearing fractions was higher, than from Y-bearing fractions. The high total motility of X- and Y-bearing spermatozoa as well as greater percent sperms with progressive motility were observed after use of BSA protocol. The application of TLR7/8 ligand R848 protocol led to reducing of Y-sperm motility and enhancement of non-progressivity in both fractions, which corresponded to the determined high amount of the extracellular lactate. For both methods, the significantly reduced activity of enzymes in the X and Y spermatozoa environmental fluids was established. Both protocols produce X- and Y-sperm fractions with satisfactory quality (over 80% total motility and over 50% rapid and medium progressive spermatozoa in each fraction).
摘要译文
性别分级精液中分离的分数的质量对于人工授精的成功非常重要。这项研究旨在评估X-和Y含毛状的Ram SpermatozoA的一些体外特征（DNA数量，运动学参数和酶活性），这些精子由牛血清白蛋白（BSA）柱和Toll-like受体（TLR）7/8配体R848 R848 R848 R848 R848分类。。通过人造阴道收集了六只公羊的射精，并进行了计算机辅助的精液分析（CASA）。分析了整个射精和X和Y级分中具有快速和中等渐进率或非促进性的精子的总运动和百分比。生物化学分析仪测量酶ALP，GGT，CK，LDH和乳酸在射精和性精子的环境流体中的积累。从沉淀的精子中分离出DNA，并测量其数量。对于这两种方案，来自X含量分数的DNA质量都高于含Y的分数。使用BSA方案后，观察到了X-和Y含X和Y含量的精子的高总运动以及具有渐进运动性的精子。TLR7/8配体R848方案的应用导致y-sperm运动的降低和在这两个级分中的非促进性增强，这对应于确定的大量细胞外乳酸。对于这两种方法，都建立了X和Y精子环境流体中酶的活性显着降低。这两种方案都产生X和Y-Sperm的质量令人满意的质量（总运动性超过80％，每分级的快速和中等渐进式精子超过50％）。
Elena Kistanova [b]. In vitro characteristics of X- and Y-bearing ram spermatozoa sorted by bovine serum albumin (BSA) column and TLR7/8 ligand R848[J]. Animal Reproduction Science, 2024,263: 107450