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New approach to assess sperm DNA fragmentation dynamics： Fine-tuning mathematical models

Journal of Animal Science and Biotechnology 2017年第3期8卷 592-600页

作者： Isabel Ortiz Jesus Dorado jane morrell Jaime Gosalvez Francisco Crespo Juan M.Jimenez Manuel Hidalgo Veterinary Reproduction Group Department of Animal Medicine and Surgery Faculty of Veterinary Medicine University of Cordoba 14071 Cordoba Spain Department of Clinical Sciences Division of Reproduction Swedish University of Agricultural Sciences Box 7054 SE-75007 Uppsala Sweden Department of Biology Universidad Aut6noma de Madrid 28049 Madrid Spain Department of Reproduction Centro Militar de Cria Caballar (FESCCR-Ministry of Defense) 05005 Avila Spain Department of Chemistry and Physics of Materials University of Salzburg Hellbrunner StraBe 34/111 A-5020 Salzburg Austria

Background： Sperm DNA fragmentation（sDF） has been proved to be an important parameter in order to predict in vitro the potential fertility of a semen sample. Colloid centrifugation could be a suitable technique to select those donkey sperm more resistant to DNA fragmentation after thawing. Previous studies have shown that to elucidate the latent damage of the DNA molecule, sDF should be assessed dynamically, where the rate of fragmentation between treatments indicates how resistant the DNA is to iatrogenic damage. The rate of fragmentation is calculated using the slope of a linear regression equation. However, it has not been studied if s DF dynamics fit this model. The objectives of this study were to evaluate the effect of different after-thawing centrifugation protocols on sperm DNA fragmentation and elucidate the most accurate mathematical model（linear regression, exponential or polynomial） for DNA fragmentation over time in frozen-thawed donkey ***： After submitting post-thaw semen samples to no centrifugation（UDC）, sperm washing（SW） or single layer centrifugation（SLC） protocols, sD F values after 6 h of incubation were significantly lower in SLC samples than in SW or *** of determination（R-2） values were significantly higher for a second order polynomial model than for linear or exponential. The highest values for acceleration of fragmentation（aSDF） were obtained for SW, fol owed by SLC and ***： SLC after thawing seems to preserve longer DNA longevity in comparison to UDC and SW. Moreover,the fine-tuning of models has shown that sDF dynamics in frozen-thawed donkey semen fit a second order polynomial model, which implies that fragmentation rate is not constant and fragmentation acceleration must be taken into account to elucidate hidden damage in the DNA molecule.

关键词： Colloid centrifugation Dynamics Fine-tuning Mathematical models Sperm DNA fragmentation

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Animal Reproduction Technologies-FuturePerspectives

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Journal of Agricultural Science and Technology(A) 2016年第5期6卷 301-313页

作者： jane morrell Patrice Humblot Department of Clinical Sciences Swedish University of Agricultural Sciences (SLU) Box 7054 SE-75007 Uppsala Sweden

Reproductive biotechnologies, such as artificial insemination, sperm selection and embryo technologies, offer possibilitiesfor animal producers to increase reproductive efficiency. There have been many significant developments in reproductivebiotechnologies over the last few decades, e.g., in sperm handling and preservation, in vitro embryo production and preservation,sexing and cloning. This review discusses some of the key changes that have occurred and explores their potential for increasing thereproductive efficiency of domestic animals in the future. As a consequence, they also offer opportunities to facilitate or accelerategenetic selection. If properly used, they may contribute to increase the sustainability of animal production. The role of epigenetics ininfluencing phenotype is also considered.

关键词： Animal reproduction technologies, livestock, genetic selection, conservation breeding, sperm, embryo production,epigenetics, future perspectives.

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Colloid Centrifugation of Semen: Applications in Assisted Reproduction

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American Journal of Analytical Chemistry 2016年第8期7卷 597-610页

作者： jane M. morrell Heriberto Rodriguez-Martinez Division of Reproduction Clinical Sciences Swedish University of Agricultural Sciences Uppsala Sweden Division of Clinical Sciences/O&G Department of Clinical and Experimental Medicine Linkoping University Linkoping Sweden

Colloid centrifugation can be used to separate heterogeneous populations of cells particularly semen samples, which typically contain mixtures of spermatozoa at different stage of maturity together with dead and dying spermatozoa and also non-sperm cells. Colloids can be used to enrich the population of spermatozoa that are viable and functional;these are the spermatozoa that are needed for fertilization in Assisted Reproduction. Sperm samples obtained after colloid centrifugation may be enriched for motile, morphologically normal spermatozoa with intact chromatin. Insemination of mares with stallion sperm samples prepared by colloid centrifugation resulted in a higher pregnancy rate per cycle than controls, confirming that the good sperm attributes observed in the laboratory were indicative of functional spermatozoa. The method does not simply enrich for viable spermatozoa by inhibiting dead or dying spermatozoa from passing through the colloid since emerging evidence shows that colloid centrifugation can select for spermatozoa with certain properties such as metabolic activity. Other reproductive cells, such as spermatids or progenitor cells, may also be purified by colloid centrifugation. This review describes the history of colloid centrifugation for selecting spermatozoa, compares colloid centrifugation with alternative techniques, and finally describes some of the applications of the technique within the animal breeding industry.

关键词： Animal Spermatozoa Purification Enrichment Selection Spermatids

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