Skip to main content Accessibility help
×
Home

Evaluation of chromatin integrity of motile bovine spermatozoa capacitated in vitro

  • Z. Reckova (a1) (a2), M. Machatkova (a3) (a1), R. Rybar (a1), J. Horakova (a1), P. Hulinska (a1) and L. Machal (a2)...

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.

Copyright

Corresponding author

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

References

Hide All
Ahmadi, A. & Soon-Chye, N.G. (1999). Fertilizing ability of DNA-damaged spermatozoa. J. Exp. Zool. 284, 696704.
Aitken, R.J. & Clarkson, J.S. (1988). Significance of reactive oxygen species and antioxidants in defining the efficacy of sperm preparation techniques. J. Androl. 9, 367–76.
Alomar, M., Mahieu, J., Verhaeghe, B., Defoin, L. & Donnay, I. (2006). Assessment of sperm quality parameters of six bulls showing different abilities to promote embryo development in vitro. Reprod. Fertil. Dev. 18, 395402.
Boe-Hansen, G.B., Averz, B., Christensen, P., Lehn-Jensen, H. & Greve, T. (2003). Sperm chromatin structure and IVF in bull with low fertilization in vivo. Theriogenology 59, 439. Abstract.
Boe-Hansen, G.B., Ersbøll, A.K., Greve, T. & Christensen, P. (2005). Increasing storage time of extended boar semen reduces sperm DNA integrity. Theriogenology 63, 2006–19.
Chohan, K.R., Griffin, J.T., Lafromboise, M., De Jonge, C.J. & Carrell, D.T. (2004). Sperm DNA damage in neat and density gradient fractions of patient and donor semen samples by different chromatin evaluation assays. Fertil. Steril. 82, S95S96.
Evenson, D.P., Larson, K.L. & Jost, L.K. (2002). The sperm chromatin structure assay: clinical use for detecting sperm DNA fragmentation in male infertility and comparisons with other techniques. J. Androl. 23, 2543.
Evenson, D.P. & Wixon, R. (2006). Clinical aspects of sperm DNA fragmentation detection and male infertility. Theriogenology 65, 979–91.
Fatehi, A.N., Bevers, M.M., Schoevers, E., Roelen, B.A.J., Colenbrander, B., Gadella, B.M. (2006). DNA damage in bovine sperm does not block fertilization and early embryonic development but induces apoptosis after first cleavages. J. Androl. 27, 176–88.
Filatov, M.V., Semenova, E.V., Vorobeva, O.A., Leonteva, O.A. & Drobchenko, E.A. (1999). Relationship between abnormal sperm chromatin packing and IVF results. Mol. Human Reprod. 5, 825–30.
Gillan, L., Evans, G. & Maxwell, W.M.C. (2005). Flow cytometric evaluation of sperm parameters in relation to fertility potential. Theriogenology 63, 445–57.
Graham, J.K., Kunze, E. & Hammerstedt, R.H. (1990). Analysis of sperm cell viability, acrosomal integrity and mitochondrial function using flow cytometry. Biol. Reprod. 43, 5564.
Hallap, T., Nagy, S., Haard, M., Jaakma, U., Johannisson, A. & Rodriguez-Martinez, H. (2005). Sperm chromatin stability in frozen-thawed semen in maintained over age in AI bulls. Theriogenology 63, 1752–63.
Katska-Ksiazkiewicz, L., Bochenek, M., Rynska, B. (2005). Effect of quality of sperm chromatin structure on in vitro production of cattle embryos. Arch. Tierz. Dummerstorf 48, 32–9.
Krzyzosiak, J., Evenson, D., Pitt, C., Jost, L., Molan, P. & Vishwanath, R. (2000). Changes in susceptibility of bovine sperm to in-situ DNA denaturation, during prolonged incubation at ambient temperature under conditions of exposure to reactive oxygen species and nuclease inhibitor. Reprod. Fertil. Dev. 12, 251–61.
Larson, K.L., De Jonge, C.J., Barnes, A.M., Jost, L.K. & Evenson, D.P. (2000). Sperm chromatin structure assay parameters as predictors of failed pregnancy following assisted reproductive techniques. Human Reprod. 15, 1717–22.
Larson-Cook, K.L., Brannian, J.D., Hansen, K.A., Kasperson, K.M., Aamold, B.S. & Evenson, D.P. (2003). Relationship between the outcomes of assisted reproductive techniques and sperm DNA fragmentation as measured by the sperm chromatin structure assay. Fertil. Steril. 80, 895902.
Lewis, S.E.M. & Aitken, R.J. (2005). DNA damage to spermatozoa has impacts on fertilization and pregnancy. Cell Tissue Res. 322, 3341.
Machatkova, M., Hanzalova, K., Horakova, J., Reckova, Z. & Hulinska, P. (2006). Collection of oocytes from donors in the growth phase of follicular development can enhance the production of bovine embryos for cryopreservation. Vet. Med-Czech. 51, 232–8.
Madrid-Bury, N., Perez-Gutierrez, J.F., Perez-Garnelo, S., Moreira, P., Sanjuanbenito, B.P., Gutierrez-Adan, A. & Martinez, J.D. (2005). Relationship between non-return rate and chromatin condensation of deep frozen bull spermatozoa. Theriogenology 64, 232–41.
Mendes, J.O.B. Jr., Burns, P.D., De La Torre-Sanchez, J.F. Seidel jr., & , G.E. (2003). Effect of heparin on cleavage rates and embryo production with four bovine sperm preparation protocols. Theriogenology 60, 331–40.
Nagy, S., Hallap, T., Johannisson, A. & Rodriguez-Martinez, H. (2004). Changes in plasma membrane and acrosome integrity of frozen-thawed bovine spermatozoa during a 4 h incubation as measured by multicolor flow cytometry. Anim. Reprod. Sci. 80, 225–35.
Parrish, J.J., Krogenaes, A. & Susko-Parrish, J.L. (1995). Effect of bovine sperm separation by either swim-up or percoll method on success of in vitro fertilization and early embryonic development. Theriogenology 44, 859–69.
Payne, J.F., Raburn, D.J., Couchman, G.M., Price, T.M., Jamison, M.G. & Walmer, D.K. (2005). Redefining the relationship between sperm deoxyribonucleic acid fragmentation as measured by the sperm chromatin structure assay and outcomes of assisted reproductive techniques. Fertil. Steril. 84, 356–64.
Pereira, R.J.T.A., Tuli, R.K., Wallenhorst, S. & Holtz, W. (1996). The effect of heparin, caffeine and calcium ionophore A 23187 on in vitro induction of the acrosome reaction in frozen-thawed bovine and caprine spermatozoa. Theriogenology 54, 185–92.
Sailer, B.L., Jost, L.K. & Evenson, D.P. (1996). Bull sperm head morphometry related to abnormal chromatin structure and fertility. Cytometry 24, 167–73.
Silva, P.F.N. & Gadella, B.M.(2006). Detection of damage in mammalian sperm cells. Theriogenology 65, 958–78.
Saleh, R.A., Agarwal, A., Nelson, D.R., Nada, E.A., El-Tonsy, M.H., Alvarez, J.G., Thomas, A.J. & Sharma, R.K. (2002). Increased sperm nuclear DNA damage in normozoospermic infertile men: a prospective study. Fertil, Steril. 78, 313–8.
Smit, M., Dohle, G.R., Hop, W.C.J., Wildhagen, M.F., Weber, R.F.A. & Romijn, C. (2007). Clinical correlates of the biological variation of sperm DNA fragmentation in infertile men attending an andrology outpatient clinic. Int. J. Androl. 30, 4855.
Spanó, M., Bonde, J.P., Hjollund, H.I., Kolstad, H.A., Cordelli, E. & Leter, G. (2000). Sperm chromatin damage impairs human fertility. Fertil. Steril. 73, 4350.
Sumantri, C., Ooe, M., Saha, S. & Boediono, A. (1996). The influence of sperm–oocyte incubation time and breed of bull on in vitro embryo development in cattle. Theriogenology 45, 264. Abstract.
Van Soom, A. & de Kruif, A. (1996). Oocyte maturation, sperm capacitacion and preimplantation development in the bovine: Implications for in vitro production of embryos. Reprod. Domestic Anim. 31, 687701.
Virant-Klun, I., Tomazevic, T. & Meden-Vrtovec, H. (2002). Sperm single-stranded DNA, detected by acridine orange staining, reduces fertilization and quality of ICSI-derived embryos. J. Assist. Reprod. Genet. 19, 319–28.
Virro, M.R., Larson-Cook, K.L. & Evenson, D.P. (2004). Sperm chromatin structure assay (SCSA) parameters are related to fertilization, blastocyst development and ongoing pregnancy in in vitro fertilization and intracytoplasmic sperm injection cycles. Fertil. Steril. 81, 1289–95.
Zalata, A., Hafez, T. & Comhaire, F. (1995). Evolution of the role of reactive oxygen species in male infertility. Human Reprod. 10, 1444–51.
Zini, A., Bielecki, R., Phang, D. & Zenzes, M.T. (2001). Correlations between two markers of sperm DNA integrity, DNA denaturation and DNA fragmentation, in fertile and infertile men. Fertil. Steril. 75, 674–7.

Keywords

Evaluation of chromatin integrity of motile bovine spermatozoa capacitated in vitro

  • Z. Reckova (a1) (a2), M. Machatkova (a3) (a1), R. Rybar (a1), J. Horakova (a1), P. Hulinska (a1) and L. Machal (a2)...

Metrics

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed