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Evaluation of chromatin integrity of motile bovine spermatozoa capacitated in vitro

Published online by Cambridge University Press:  01 August 2008

Z. Reckova ,
M. Machatkova ,
R. Rybar ,
J. Horakova ,
P. Hulinska  and
L. Machal
Z. Reckova
Affiliation:
Department of Genetics and Reproduction, Veterinary Research Institute, Hudcova 70, 621 00 Brno, Czech Republic. Department of Animal Breeding, Mendel University of Farming and Forestry, Zemedelska 1, 613 00 Brno, Czech Republic.
M. Machatkova*
Affiliation:
Department of Genetics and Reproduction, Veterinary Research Institute, Hudcova 70, 621 00 Brno, Czech Republic. Department of Genetics and Reproduction, Veterinary Research Institute, Hudcova 70, 621 00 Brno, Czech Republic.
R. Rybar
Affiliation:
Department of Genetics and Reproduction, Veterinary Research Institute, Hudcova 70, 621 00 Brno, Czech Republic.
J. Horakova
Affiliation:
Department of Genetics and Reproduction, Veterinary Research Institute, Hudcova 70, 621 00 Brno, Czech Republic.
P. Hulinska
Affiliation:
Department of Genetics and Reproduction, Veterinary Research Institute, Hudcova 70, 621 00 Brno, Czech Republic.
L. Machal
Affiliation:
Department of Animal Breeding, Mendel University of Farming and Forestry, Zemedelska 1, 613 00 Brno, Czech Republic.
*
All correspondence to: Marie Machatkova. Department of Genetics and Reproduction, Veterinary Research Institute, Hudcova 70, 621 00 Brno, Czech Republic. Tel: +420 533 331 418. Fax: +420 541 211 229. e-mail: machatkova@vri.cz
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Summary

The efficiency of in vitro embryo production is highly variable amongst individual sires in cattle. To eliminate that this variability is not caused by sperm chromatin damage caused by separation or capacitacion, chromatin integrity was evaluated. Seventeen of AI bulls with good NRRs but variable embryo production efficiency were used. For each bull, motile spermatozoa were separated on a Percoll gradient, resuspended in IVF–TALP medium and capacitated with or incubated without heparin for 6 h. Samples before and after separation and after 3-h and 6-h capacitacion or incubation were evaluated by the Sperm Chromatin Structure Assay (SCSA) and the proportion of sperm with intact chromatin structure was calculated. Based on changes in the non-DFI-sperm proportion, the sires were categorized as DNA-unstable (DNA-us), DNA-stable (DNA-s) and DNA-most stable (DNA-ms) bulls (n = 3, n = 5 and n = 9, respectively). In DNA-us bulls, separation produced a significant increase of the mean non-DFI-sperm proportion (p ≤ 0.01), as compared with the value before separation. Capacitacion produced a significant decrease in the mean non-DFI-sperm proportion in H+ sperm (p ≤ 0.01). In DNA-s bulls, separation significantly increased the mean non-DFI-sperm proportion (p ≤ 0.01) but during capacitacion, the mean non-DFI-sperm proportion remained almost unchanged. In DNA-ms bulls, neither separation nor capacitacion had any effect on the mean non-DFI-sperm proportion. It can be concluded that, although separation and capacitacion may produce some changes in sperm chromatin integrity, these are not associated with different in vitro fertility of the bulls involved.

Keywords

BullsChromatin integrityIn vitro capacitacionSpermatozoa
Type
Research Article
Information
Zygote , Volume 16 , Issue 3 , August 2008 , pp. 195 - 202
Copyright
Copyright © Cambridge University Press 2008

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