The objectives of this study were to determine what effects iodixanol would have on total and progressive motility, plasma membrane integrity (viability), acrosome integrity and DNA structure when present during cryopreservation of equine spermatozoa,. We hypothesized that the addition of iodixanol would improve post-thaw values for measured parameters. Ejaculates from six stallions were collected, centrifuged at 900 x g for ten minutes to remove supernatant, and suspended to 200 x 106 cells/ml with 0%, 2.5% and 5% iodixanol in an egg-yolk based extender and cryopreserved. Before and after cryopreservation sperm motility was assessed by computer assisted semen analysis, and samples were stained with SYBR-14/propidium iodide (PI) for viability, with PI/fluorescent isothiocynate-PNA (Arachis Hypogaea) for acrosome integrity and assessed by flow cytometry. Sperm DNA was evaluated using the sperm chromatin structure assay test and assessed by flow cytometry. The mean (± S.E.) percentage pre- and post-thaw total motility, progressive motility, viability, acrosome reactivity, COMPát and MEANát were analyzed using Shapiro-Wilk test to evaluate if data followed a normal distribution. When results followed a normal distribution an ANOVA was performed and where a significant interaction of treatment was observed (p=0.05), a Tukey-Kramer post-hoc test was applied. If results did not follow a normal distribution, a binomial logistical regression was performed. The 5% post-thaw treatment group showed increased viability and decreased DNA damage (p<0.001). Although the 0% group showed greater total and progressive motility than the 2.5% group, it was not greater than the 5% group. The 5% post-thaw treatment group had significantly more (p<0.001) non-reacted and damaged acrosomes than both the 0% and 2.5% groups. These findings suggest that the presence of iodixanol during cryopreservation may have a beneficial effect by protecting plasma membrane and sperm DNA, but the exact mechanism of action is unknown.
| Identifer | oai:union.ndltd.org:LSU/oai:etd.lsu.edu:etd-11142012-084124 |
| Date | 26 November 2012 |
| Creators | Beehan, David Paul |
| Contributors | Lyle, Sara K., Eilts, Bruce E., Paccamonti, Dale L. |
| Publisher | LSU |
| Source Sets | Louisiana State University |
| Language | English |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | http://etd.lsu.edu/docs/available/etd-11142012-084124/ |
| Rights | restricted, I hereby certify that, if appropriate, I have obtained and attached herein a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted is the same as that approved by my advisory committee. I hereby grant to LSU or its agents the non-exclusive license to archive and make accessible, under the conditions specified below and in appropriate University policies, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report. |
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