Kromatinstabilitet som grund för kvalitetsbedömning av hingstsperma
This study is part of a project to evaluate and develop metods to control the semen quality in stallions. The study was carried out in cooperation between SLU and Flyinge AB. The objective of the study was to analyse -if there are individual differences in sperm chromatin integrety between differe...
| Autor principal: | |
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| Formato: | L3 |
| Lenguaje: | sueco Inglés |
| Publicado: |
SLU/Dept. of Clinical Sciences (until 231231)
2007
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| Materias: |
| _version_ | 1855571920146137088 |
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| author | Hammar, Linda |
| author_browse | Hammar, Linda |
| author_facet | Hammar, Linda |
| author_sort | Hammar, Linda |
| collection | Epsilon Archive for Student Projects |
| description | This study is part of a project to evaluate and develop metods to control the semen quality in stallions. The study was carried out in cooperation between SLU and Flyinge AB. The objective of the study was to analyse
-if there are individual differences in sperm chromatin integrety between different stallions
-if there is a correlation between subjectively evaluated sperm motility and sperm chromatin integrity
-if sperm chromatin integrity is changed after selection of the semen with a centrifugation method through the silica-spheres, gradient or a single-layer, i.e to use the chromatin integrity analysis as a method to evaluate the selection method.
The use of chilled transported semen has increased a lot during the last 10 years, especially among the Swedish warmblood breed. Because the foaling percentage has decreased with the increased use of chilled transported semen, there is a need to develop methods to control the sperm quality. Evaluation of the chromatin integrity could be one way to evaluate the quality of the semen.
Chromatin is the DNA in the nucleus of the sperm. If the chromatin is denatured it may lead to a failed fertilization or a disturbed development of the embryo or the placenta.
Ten stallions of Swedish Warmblood breed at the Flyinge National Stud with the age 4-23 years were used in the study. The stallion semen was collected at seven times during a period of 2 ½ weeks. From every ejaculate samples were taken both before and after centrifugation through either of two different silica-spheres, single-layer or gradient for chromatin analysis and evaluation of sperm motility. The samples were immediatly frozen in TNE buffer and transported to Uppsala where they were analyzed with help of flow-cytometry. The results was presented as DFI (DNA Fragmentation Index) which measures the percentage of the sperms fragmentated DNA, and also the main DFI (X-DFI) and the standard deviation of DFI (SD-DFI).
The result showed that the method of sperm centrifugation was effective in the selection of sperms with damage chromatin integrity. There was no difference in effectiveness between the two different metods of centrifugation, single–layer and gradient.
The DNA Fragmentation Index (DFI) and the subjectively evaluated sperm motility was negative correlated. This means that the lower the DFI, the higher the motility. The result also showed that there was a statistical significant difference in DFI between stallions. |
| format | L3 |
| id | RepoSLU11691 |
| institution | Swedish University of Agricultural Sciences |
| language | swe Inglés |
| publishDate | 2007 |
| publishDateSort | 2007 |
| publisher | SLU/Dept. of Clinical Sciences (until 231231) |
| publisherStr | SLU/Dept. of Clinical Sciences (until 231231) |
| record_format | eprints |
| spelling | RepoSLU116912017-11-23T13:34:55Z Kromatinstabilitet som grund för kvalitetsbedömning av hingstsperma Hammar, Linda kromatinstabilitet kvalitetsbedömning hingstsperma flödescytometri centrifugeringsmetod selektionsmetod This study is part of a project to evaluate and develop metods to control the semen quality in stallions. The study was carried out in cooperation between SLU and Flyinge AB. The objective of the study was to analyse -if there are individual differences in sperm chromatin integrety between different stallions -if there is a correlation between subjectively evaluated sperm motility and sperm chromatin integrity -if sperm chromatin integrity is changed after selection of the semen with a centrifugation method through the silica-spheres, gradient or a single-layer, i.e to use the chromatin integrity analysis as a method to evaluate the selection method. The use of chilled transported semen has increased a lot during the last 10 years, especially among the Swedish warmblood breed. Because the foaling percentage has decreased with the increased use of chilled transported semen, there is a need to develop methods to control the sperm quality. Evaluation of the chromatin integrity could be one way to evaluate the quality of the semen. Chromatin is the DNA in the nucleus of the sperm. If the chromatin is denatured it may lead to a failed fertilization or a disturbed development of the embryo or the placenta. Ten stallions of Swedish Warmblood breed at the Flyinge National Stud with the age 4-23 years were used in the study. The stallion semen was collected at seven times during a period of 2 ½ weeks. From every ejaculate samples were taken both before and after centrifugation through either of two different silica-spheres, single-layer or gradient for chromatin analysis and evaluation of sperm motility. The samples were immediatly frozen in TNE buffer and transported to Uppsala where they were analyzed with help of flow-cytometry. The results was presented as DFI (DNA Fragmentation Index) which measures the percentage of the sperms fragmentated DNA, and also the main DFI (X-DFI) and the standard deviation of DFI (SD-DFI). The result showed that the method of sperm centrifugation was effective in the selection of sperms with damage chromatin integrity. There was no difference in effectiveness between the two different metods of centrifugation, single–layer and gradient. The DNA Fragmentation Index (DFI) and the subjectively evaluated sperm motility was negative correlated. This means that the lower the DFI, the higher the motility. The result also showed that there was a statistical significant difference in DFI between stallions. Denna studie ingår i ett projekt som har som mål är att utvärdera och vidareutveckla metoder för kvalitetsbedömning av hingstsperma avsedd för semin. Studien har genomförts i samarbete mellan SLU och Flyinge AB. Syftet med studien var att undersöka -om det finns individuella skillnader mellan hingstar vad gäller spermiernas kromatinstabilitet -om det finns korrelation mellan subjektivt bedömd spermiemotilitet och kromatinstabilitet hos spermier -om kromatinstabilitet hos spermier förändras efter centrifugering genom single layer och gradient dvs använda kromatinanalys som ett mått på om selektionsmetoden fungerar Under de senaste 10 åren har det skett en kraftig ökning av användningen av kyld transportsperma ffa inom svenskvarmblodsavel. Eftersom fölningsprocenten sjunkit i takt med att verksamheten med kyld sperma ökat är det av stort intresse att utveckla metoder för att kvalitetsbedöma hingstsperma. En bedömning av kromatinets kvalitet skulle kunna utgöra en kvalitetskontroll av hingstsperma för semin. Kromatinet finns i spermiekärnan. Skador i kromatinet kan leda till en misslyckad befruktning då spermiens kärna inte dekondenseras normalt och det befruktade ägget dör eller till en felaktig embryoutveckling. Tio hingstar av halvblodsras stationerade i Flyinge i åldrarna 4-23 år ingick i studien. Hingstarna samlades på sperma vid 7 tillfällen under en period på 2 ½ vecka. För varje spermaejakulat togs prover för kromatinundersökning samt bedömdes motiliteten, både före centrifugering och efter centrifugering av ejakulaten genom två olika silikatsfärer, ”Single layer” och ”Gradient”. Spermieproverna frystes omedelbart in med TNE buffert i flytande kväve och transporterades sedan till Uppsala där de analyserades med hjälp av flödescytometri. Resultaten presenterades som DFI (DNA Fragmentation Index) vilket är ett mått på hur stor andel av spermierna som har skadat DNA, angivet i procent, samt X-DFI (medelvärdet för DFI) och SD-DFI (standardavvikelsen för DFI). Resultaten i denna studie visade att selektionsmetoden var effektiv när det gällde att selektera bort spermier med skadat kromatin. Däremot skiljde sig inte de två selektionsmetoderna med gradient centrifugering och single layer centrifugering åt i effektivitet när de jämfördes med varandra. Resultaten visade också att det fanns en statistiskt signifikant skillnad i DFI mellan hingstar vad gäller kromatinstabilitet samt en negativ korrelation mellan DFI och motilitet. Ju lägre DFI (dvs låg andel spermier med skadat kromatin) desto högre motilitet. SLU/Dept. of Clinical Sciences (until 231231) 2007 L3 swe eng https://stud.epsilon.slu.se/11691/ |
| spellingShingle | kromatinstabilitet kvalitetsbedömning hingstsperma flödescytometri centrifugeringsmetod selektionsmetod Hammar, Linda Kromatinstabilitet som grund för kvalitetsbedömning av hingstsperma |
| title | Kromatinstabilitet som grund för kvalitetsbedömning av hingstsperma |
| title_full | Kromatinstabilitet som grund för kvalitetsbedömning av hingstsperma |
| title_fullStr | Kromatinstabilitet som grund för kvalitetsbedömning av hingstsperma |
| title_full_unstemmed | Kromatinstabilitet som grund för kvalitetsbedömning av hingstsperma |
| title_short | Kromatinstabilitet som grund för kvalitetsbedömning av hingstsperma |
| title_sort | kromatinstabilitet som grund för kvalitetsbedömning av hingstsperma |
| topic | kromatinstabilitet kvalitetsbedömning hingstsperma flödescytometri centrifugeringsmetod selektionsmetod |