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Viability and apoptosis in spermatozoa of transgenic rabbits

Published online by Cambridge University Press:  13 December 2010

P. Chrenek ,
A.V. Makarevich  and
M. Simon
P. Chrenek*
Affiliation:
Animal Production Research Centre Nitra, 95141 Luzianky near Nitra, Slovak Republic. Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture, Nitra, Slovak Republic.
A.V. Makarevich
Affiliation:
Animal Production Research Centre Nitra, Slovak Republic.
M. Simon
Affiliation:
Institute of Animal Biochemistry and Genetics, Slovak Academy Science, Ivanka pri Dunaji, Slovak Republic.
*
All correspondence to: P. Chrenek. Animal Production Research Centre Nitra, 95141 Luzianky near Nitra, Slovak Republic. Tel: + 421 37 6546 289. Fax: + 421 37 6546 285. e-mail: chrenekp@yahoo.com
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Summary

The aim of our study was to compare the viability of sperm cells from transgenic (mWAP-hFVIII gene) or non-transgenic (normal) rabbit males as assessed by viability (SYBR-14/PI) and apoptosis (annexin V) tests. These results were evaluated using female conception rates following insemination with the respective sperm samples. No significant differences were found in concentration and motility between transgenic and non-transgenic spermatozoa. Spermatozoa from both transgenic (63.05 ± 20.05%) or non-transgenic (65.75 ± 22.15%) males, stained with SYBR-14 (green), were found to be morphologically normal. In both groups, the highest proportion of annexin V-positive sperm staining was found in the post-acrosomal part of the sperm head (8.66 and 27.53%). The percentage of sperm that stained with SYBR-14/PI or with annexin V/DAPI was correlated with liveborn in transgenic rabbits (R2 = 0.6118 and R2 = 0.2187, respectively) or non-transgenic rabbits (R2 = 0.671 and R2 = 0.3579, respectively). These data indicate that there was no difference in the viability of rabbit transgenic and non-transgenic spermatozoa when determined by both fluorescence assays.

Keywords

Annexin VRabbitSpermatozoaSYBR-14Transgenic
Type
Research Article
Information
Zygote , Volume 20 , Issue 1 , February 2012 , pp. 33 - 37
Copyright
Copyright © Cambridge University Press 2010

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References

Bialkowska, J., Demianowicz, W. & Glogowski, J. (2004). Determining the viability of zebra mussel (Dreissena polymorpha) spermatozoa and changes in the integrity of its membrane using the fluorescence method. Arch. Polish Fish. 12, 23–30.Google Scholar
Chrenek, P., Vasicek, D., Makarevich, A., Jurcik, R., Suvegova, K., Bauer, M., Parkanyi, V., Rafay, J., Batorova, A. & Paleyanda, R.K. (2005). Increased transgene integration efficiency upon microinjection of DNA into both pronuclei of rabbit embryos. Transgenic Res. 14, 417–28.CrossRefGoogle ScholarPubMed
Chrenek, P., Dragin, S. & Makarevich, A.V. (2006). Reproductive characteristics of transgenic rabbit males with human protein C gene. Slovak J. Anim. Sci. 39, 131–4.Google Scholar
Chrenek, P., Trandzik, J., Massanyi, P., Makarevich, A., Lukac, N., Peskovicova, D. & Paleyanda, R.K. (2007a). Effect of transgenesis on reproductive traits of rabbit males. Anim. Reprod. Sci. 99, 127–34.CrossRefGoogle ScholarPubMed
Chrenek, P., Massányi, P., Makarevich, A.V., Pukač, N., Zahradnikova, M., Schneidgenova, M. & Ryban, L. (2007b). Rabbit transgenic brothers with different reproductive traits. Slovak J. Anim. Sci. 40, 113–7.Google Scholar
Gadella, B.M. & Harrison, R.A.P. (2002). Capacitation induces cyclic adenosine 3′,5′-monophosphate-dependent, but apoptosis-unrelated, exposure of aminophospholipids at the apical head plasma membrane of boar sperm cells. Biol. Reprod. 67, 1340–50.CrossRefGoogle ScholarPubMed
Garner, D.L. & Johnson, L. (1995). Viability assessment of mammalian spermatozoa using SYBR-14 and propidium iodide. Biol. Reprod. 53, 276–84.CrossRefGoogle ScholarPubMed
Garner, D.L., Johnson, L.A., Allen, C.H., Palencia, D.D. & Chambers, C.S. (1996). Comparison of seminal quality in Holstein bull as yearling and as mature sires. Theriogenology 45, 923–34.CrossRefGoogle Scholar
Harrison, R.A. & Vickers, S.E. (1990). Use of fluorescent probes to assess membrane integrity in mammalian spermatozoa. J. Reprod. Fertil. 88, 343–52.CrossRefGoogle ScholarPubMed
Hong, C.Y., Juany, J.J., Wu, P., Lo, S. & Wei, Y.H. (1988). Fluorescence supravita stain human sperm correlation with sperm motility measured by a transmembrane migration metod. Andrologia 20, 516–20.CrossRefGoogle Scholar
Januskauskas, A., Johannisson, A. & Rodriguez-Martinez, H. (2003). Sibyle membrane changes in cryopreserved bull semen in relation with sperm viability chromatin structure, and field fertility. Theriogenology 60, 743–58.CrossRefGoogle ScholarPubMed
Makarevich, A.V., Parkanyi, V., Ondruška, Ľ., Kubovičová, E., Fľak, P., Slezáková, M., Pivko, J. & Rafay, J. (2008a). Evaluation of fertilizing capacity of rabbit sperm on the basis of annexin V-labelled membrane changes. Slovak J. Anim. Sci. 41, 1–5.Google Scholar
Makarevich, A.V., Chrenek, P., Massányi, P., Lukac, N. & Pivko, J. (2008b). Apoptosis as a tool for determination of animal embryo quality. Slovak J. Anim. Sci. 41, 153–9.Google Scholar
Martin, G., Sabido, O., Durand, P. & Levy, R. (2004). Cryopreservation induces an apoptosis-like mechanism in bull sperm. Biol. Reprod. 71, 28–37.CrossRefGoogle Scholar
Muratori, M., Porazzi, I., Luconi, M., Marchiani, S., Forti, G. & Baldi, E. (2004). Annexin V binding and merocyanine staining fail to detect human sperm capacitation. J. Androl. 25, 797–810.CrossRefGoogle ScholarPubMed
Pena, F.J., Johannisson, A., Wallgren, M. & Rodriguez-Martinez, H. (2003). Assessment of fresh and frozen–thawed boar semen using an annexin-V assay: a new method of evaluating sperm membrane integrity. Theriogenology 60, 677–89.CrossRefGoogle ScholarPubMed
Vermes, I., Haanen, C., Steffens-Nakken, H. & Reutelingsperger, C.P.M. (1995). A novel assay for apoptosis: flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled annexin V. J. Immunol. Meth. 184, 39–51.CrossRefGoogle ScholarPubMed
Vetter, C.M., Millet, J.E., Crawford, L.M., Armstrong, M.J., Clair, J. H., Conner, M.W., Wise, L.D. & Skopek, T.R. (1998). Comparison motility and membrane integrity to assess sperm viability. Reprod. Toxicol. 12, 105–14.CrossRefGoogle Scholar