Departamento de Reproducción Animal, INIA, Madrid, Spain.
Departamento de Mejora Genética Animal, INIA, Madrid, Spain.
PLoS One. 2019 Jan 4;14(1):e0209910. doi: 10.1371/journal.pone.0209910. eCollection 2019.
Seminal plasma is a key biological fluid that modulates sperm function in the reproduction process. However, its role in sperm biotechnologies is scarce in poultry. The aims of the present study were to study the amino acids profile and total proteins of seminal plasma in 12 Spanish chicken breeds and to investigate the role of seminal plasma on cryoresistance of rooster sperm. To investigate the role of seminal plasma on cryoresistance, diluted pooled semen samples were cryopreserved in the presence and absence of seminal plasma. Glutamic acid was the most abundant free amino acid in seminal plasma, followed by alanine, serine, valine, and glycine. There was an influence of breed (P<0.05) on the percentage of viable sperm after freezing-thawing of samples with seminal plasma. Cluster analysis revealed that White Prat, Black Castellana, Blue Andaluza, Quail Castellana, and Red-Barred Vasca returned the best freezing-thawing response (good freezers). There was a positive correlation between seminal plasma concentrations of valine, isoleucine lysine, leucine and post thaw viability. The evaluation of fertilization capacity of frozen-thawed semen from the breeds White Prat ('good freezer') and Black-Red Andaluza ('bad freezer') showed that good freezer had higher fertility (20/68, 29.4%) compared to bad freezer breed (14/76, 18.4%), even if the difference was not significant (P = 0.08). The TUNEL assay revealed that freezing/thawing procedures in presence of seminal plasma provoked higher DNA fragmentation in most of the breeds, with a positive correlation between seminal alanine, valine, isoleucine, methionine, leucine, tyrosine, phenylalanine concentrations and DNA integrity. DNA fragmentation was lower in absence of seminal plasma and the breed effect on sperm viability was highly reduced. It is concluded that specific seminal plasma amino acids were associated with post-thaw percentage of viable sperm and DNA integrity. The removal of seminal plasma decreases the variability of the results and DNA fragmentation damages.
精浆是一种调节生殖过程中精子功能的关键生物液。然而，其在禽类精子生物技术中的作用尚不清楚。本研究旨在研究 12 个西班牙鸡品种的精浆氨基酸谱和总蛋白，并探讨精浆对公鸡精子抗冷冻能力的作用。为了研究精浆对抗冷冻能力的作用，在有或没有精浆的情况下，将稀释的混合精液样本冷冻保存。精浆中最丰富的游离氨基酸是谷氨酸，其次是丙氨酸、丝氨酸、缬氨酸和甘氨酸。在有精浆的情况下，冷冻解冻样品后存活精子的百分比受品种（P<0.05）的影响。聚类分析显示，White Prat、Black Castellana、Blue Andaluza、Quail Castellana 和 Red-Barred Vasca 具有最佳的冷冻解冻反应（良好的冷冻剂）。精浆中缬氨酸、异亮氨酸、赖氨酸、亮氨酸的浓度与解冻后活力呈正相关。对 White Prat（良好冷冻剂）和 Black-Red Andaluza（不良冷冻剂）两个品种的冷冻解冻精液的受精能力进行评估，结果显示，良好冷冻剂的受精率（20/68，29.4%）高于不良冷冻剂（14/76，18.4%），尽管差异无统计学意义（P = 0.08）。TUNEL 试验表明，在有精浆的情况下，冷冻/解冻过程会导致大多数品种的 DNA 碎片化增加，精浆中丙氨酸、缬氨酸、异亮氨酸、蛋氨酸、亮氨酸、酪氨酸、苯丙氨酸浓度与 DNA 完整性呈正相关。在没有精浆的情况下，DNA 碎片化减少，品种对精子活力的影响大大降低。因此，特定的精浆氨基酸与解冻后精子的存活率和 DNA 完整性有关。去除精浆可减少结果的变异性和 DNA 碎片化损伤。
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