Relationship of seminal plasma level and extender type to sperm motility and DNA integrity. Theriogenology

Department of Physiology and Pharmacology, Texas A and M University College of Veterinary Medicine, Collage Station, TX 77843-4475, USA.
Theriogenology (Impact Factor: 1.8). 04/2005; 63(6):1584-91. DOI: 10.1016/j.theriogenology.2004.05.030
Source: PubMed


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.

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    • "Previous research has consistently demonstrated heat loads cause loss of cell function associated with chromatin damage. However, two recent studies [9] [10] dispute the association. These conflicting findings raise doubt as to the utility of measuring chromatin damage as a measure of stallion fertility and if such damage could be caused by ambient temperatures. "

    Journal of Equine Veterinary Science 09/2015; DOI:10.1016/j.jevs.2015.09.005 · 0.87 Impact Factor
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    • "The integrity of sperm DNA has a vital importance to the sperm cell. Some authors suggest that sperm DNA integrity is a more objective marker of sperm function as opposed to the sperm parameters such as motility [20]. DNA damage could be result of free radical induced damage [28] [39]. "
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    ABSTRACT: This study evaluated the protective effects of supplementation with three different sugars on the motility, morphology and DNA integrity of rat epididymal sperm chilled and stored at 4°C Epididymides were obtained from each donor. Rat epididymal sperm was diluted in Ham's F10 plus raffinose, Ham's F10 plus trehalose, Ham's F10 plus fructose, and Ham's F10 medium for control purposes. Thereafter, the extended sperm were chilled and stored in liquid form at 4°C. Sperm motility, morphological abnormalities and DNA damage were determined at 0 and 12h after chilling. No significant difference was observed in any of the parameters evaluated at 0h, before storage (P>0.05). After 12h of storage, all sugar additives led to statistically higher motility, normal sperm morphology and DNA integrity in comparison to the control group. Raffinose gave the best motility percentages (32.86±1.84%) after 12h of storage at 4°C, compared to the other groups (P<0.001). In conclusion, Raffinose, trehalose and fructose provided a better protection of sperm functional parameters against chilling injury, in comparison to the control group.
    Cryobiology 05/2012; 65(2):93-7. DOI:10.1016/j.cryobiol.2012.05.007 · 1.59 Impact Factor
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    • "In other hand, Ball et al. (2000) found that removal of seminal plasma increased the susceptibility of stallion spermatozoa to oxidative stress. This result agreed with Love et al. (2005) where found that centrifugation and removal of the seminal plasma may enhance sperm motility of cooled-stored stallion semen, but also had an adverse effect on sperm DNA integrity. "
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    ABSTRACT: Fresh semen samples were collected from 11 warm blood stallions, each ejaculate was distributed into three equal parts. The first part was diluted in a skim milk-glucose diluent (SMG), the second part was diluted in a skim milk-glucose supplemented with Tyrode's medium (SMG-T), the third part was centrifuged to remove the seminal plasma, then the sperm was resuspended in the second diluent (SMG-T-C). The diluted semen were evaluated immediately after dilution (0 hour) and at 24, 48, 72, and 96 hours of storage at 5°C. Flow cytometry was performed to determine sperm viability, mitochondrial activity and acrosomal integrity. Immediately after dilution the tested parameters of sperms that diluted in SMG-T was significantly (P<0.001) higher than those diluted with SMG and SMG-T-C, and with SMG-T-C were higher significantly (P<0.05) than those diluted with SMG. The decreasing rate in tested sperm parameter was greater significantly (P<0.001) in semen samples which were diluted with SMG than those diluted with SMG-T and SMG-T-C. In conclusion, the present study indicated that viability, acrosomal integrity, and mitochondrial activity of stallion sperms were better preserved in SMG-T in comparison with SMG, also centrifugation and removal of the seminal plasma have an adverse effect on these three sperm parameters.
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