Love C C, Brinsko S P, Rigby S L, Thompson J A, Blanchard T L, Varner D D
Department of Physiology and Pharmacology, Texas A and M University College of Veterinary Medicine, Collage Station, TX 77843-4475, USA.
Theriogenology. 2005 Apr 1;63(6):1584-91. doi: 10.1016/j.theriogenology.2004.05.030.
The relationship between seminal plasma level (0, 10, or 20%) and extender type [Kenney type (EZ-Mixin-CST) or Kenney-modified Tyrodes-KMT] to the susceptibility of sperm DNA to denaturation and sperm motility measures were investigated in cooled (5 degrees C) stallion sperm. Three ejaculates from each of three fertile stallions were collected in an artificial vagina and processed as follows: diluted one part uncentrifuged semen with four parts of extender to a final concentration of 20% seminal plasma in either CST or KMT (20% CST; 20% KMT); diluted to a final concentration of 25 million sperm/mL in either CST or KMT (10% CST; 10% KMT); centrifuged to remove virtually all seminal plasma and resuspended in either CST or KMT (0% CST-Cent; 0% KMT-Cent); centrifuged semen to remove virtually all seminal plasma and resuspended with previously filtered seminal plasma from the same stallion in either CST or KMT to a final concentration of 20% seminal plasma (20% CST-Cent; 20% KMT-Cent). Sperm motion characteristics were determined by CASA and DNA integrity (%COMP, percent of cells outside the main population) evaluated by the Sperm Chromatin Structure Assay prior to cooling, and after 24 and 48 h cooled-storage at 5 degrees C. After 48 h of storage at 5 degrees C, extenders with 0% seminal plasma (0% CST-Cent, 0% KMT-Cent) maintained highest quality DNA (P < 0.05), but 0% KMT-Cent maintained higher velocity measures (P < 0.05) than 0% CST-Cent. Total sperm motility was highest (P < 0.05) in 0% CST-Cent, 0% KMT-Cent, 10% CST, 20% CST-Cent, and 20% CST compared to the other treatment groups. Progressive sperm motility was highest (P < 0.05) after 48 h of storage in the treatment with 10% seminal plasma in Kenney extender (10% CST), despite a reduction in DNA integrity. Regardless of extender type, addition of 20% seminal plasma following centrifugation resulted in almost a two-fold increase in %COMP(alpha t), even though one of the treatments (20% CST-Cent) maintained total and progressive motility similar to treatments with no seminal plasma, suggesting that sperm motility and DNA integrity may respond independently to environmental conditions. Overall, better quality sperm features (motility and DNA) were maintained in sperm from which seminal plasma was removed followed by resuspension in either Kenney extender or modified Kenney Tyrodes-type extender.
研究了在冷却（5摄氏度）的种马精子中，精浆水平（0%、10%或20%）和稀释液类型[肯尼氏型（EZ-Mixin-CST）或改良肯尼氏泰罗德氏液-KMT]对精子DNA变性敏感性及精子活力指标的影响。从三匹生育能力正常的种马中，每匹种马采集三份精液，通过人工阴道采集后按如下方法处理：将一份未离心的精液与四份稀释液混合，使最终精浆浓度在CST或KMT中达到20%（20% CST；20% KMT）；在CST或KMT中稀释至最终浓度为2500万精子/毫升（10% CST；10% KMT）；离心以去除几乎所有精浆，然后在CST或KMT中重悬（0% CST-Cent；0% KMT-Cent）；将精液离心以去除几乎所有精浆，然后与同一匹种马先前过滤的精浆在CST或KMT中重悬，使最终精浆浓度达到20%（20% CST-Cent；20% KMT-Cent）。在冷却前、以及在5摄氏度下冷却储存24小时和48小时后，通过计算机辅助精子分析（CASA）测定精子运动特征，并通过精子染色质结构分析评估DNA完整性（%COMP，主要群体外细胞的百分比）。在5摄氏度储存48小时后，精浆为0%的稀释液（0% CST-Cent，0% KMT-Cent）维持了最高质量的DNA（P<0.05），但0% KMT-Cent比0% CST-Cent维持了更高的速度指标（P<0.05）。与其他处理组相比，0% CST-Cent、0% KMT-Cent、10% CST、20% CST-Cent和20% CST中的总精子活力最高（P<0.05）。尽管DNA完整性有所下降，但在肯尼氏稀释液中含有10%精浆的处理组（10% CST）储存48小时后，渐进性精子活力最高（P<0.05）。无论稀释液类型如何，离心后添加20%精浆会使%COMP(αt)几乎增加两倍，尽管其中一种处理（20% CST-Cent）维持的总活力和渐进性活力与无精浆处理相似，这表明精子活力和DNA完整性可能对环境条件有独立反应。总体而言，去除精浆后再在肯尼氏稀释液或改良肯尼氏泰罗德氏型稀释液中重悬的精子，能维持更好的精子质量特征（活力和DNA）。
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