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Bovine sperm cells viability during incubation with or without exogenous DNA

Published online by Cambridge University Press:  16 June 2009

Weber Beringui Feitosa ,
Marcella Pecora Milazzotto ,
Renata Simões ,
Mariana Rovegno ,
Alessandra Coralo Nicacio ,
Aníbal Ballarotti Nascimento ,
José Sérgio Arruda Gonçalves ,
José Antonio Visintin  and
Mayra Elena Ortiz D'Ávila Assumpção
Weber Beringui Feitosa
Affiliation:
Department of Animal Reproduction, College of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil.
Marcella Pecora Milazzotto
Affiliation:
Department of Animal Reproduction, College of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil. Center of Natural Human Science, Federal University of ABC, Santo Andre, Brazil.
Renata Simões
Affiliation:
Department of Animal Reproduction, College of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil.
Mariana Rovegno
Affiliation:
Department of Animal Reproduction, College of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil.
Alessandra Coralo Nicacio
Affiliation:
Department of Animal Reproduction, College of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil. Veterinary Medicine, State University of Maranhão, São Luís, Brazil.
Aníbal Ballarotti Nascimento
Affiliation:
Department of Animal Reproduction, College of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil. Department of Dairy Science, Animal Sciences, University of Wisconsin, Madison, USA.
José Sérgio Arruda Gonçalves
Affiliation:
Department of Animal Reproduction, College of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil.
José Antonio Visintin
Affiliation:
Department of Animal Reproduction, College of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil.
Mayra Elena Ortiz D'Ávila Assumpção*
Affiliation:
Department of Animal Reproduction, College of Veterinary Medicine and Animal Science, University of São Paulo, Av. Prof. Dr. Orlando Marques de Paiva, 87–Cidade Universitária–05508–900–São Paulo, SP, Brazil. Department of Animal Reproduction, College of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil.
*
All correspondence to: Mayra Elena Ortiz D'Ávila Assumpção. Department of Animal Reproduction, College of Veterinary Medicine and Animal Science, University of São Paulo, Av. Prof. Dr. Orlando Marques de Paiva, 87–Cidade Universitária–05508–900–São Paulo, SP, Brazil. Tel: +55 11 3091 7665. Fax: +55 11 3091 7412. e-mail: meoaa@usp.br
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Summary

The aim of this study was to assess the effect of exogenous DNA and incubation time on the viability of bovine sperm. Sperm were incubated at a concentration of 5 × 106/ml with or without plasmid pEYFP–NUC. Fluorescent probes, propidium iodide/Hoechst 33342, FITC–PSA and JC-1, were used to assess plasma membrane integrity (PMI), acrosome membrane integrity (AMI) and mitochondrial membrane potential (MMP) respectively at 0, 1, 2, 3 and 4 h of incubation. Exogenous DNA addition did not affect sperm viability; however, incubation time was related to sperm deterioration. Simultaneous assessment of PMI, AMI and MMP showed a reduction in the number of sperm with higher viability (integrity of plasma and acrosome membranes and high mitochondrial membrane potential) from 58.7% at 0 h to 7.5% after 4 h of incubation. Lower viability sperm (damaged plasma and acrosome membranes and low mitochondrial membrane potential) increased from 4.6% at 0 h to 25.9% after 4 h of incubation. When PMI, AMI and MMP were assessed separately we noticed a reduction in plasma and acrosome membrane integrity and mitochondrial membrane potential throughout the incubation period. Therefore, exogenous DNA addition does not affect sperm viability, but the viability is reduced by incubation time.

Keywords

Acrosomal membraneExogenous DNAFluorescent probesMitochondrial membrane potentialPlasma membrane
Type
Research Article
Information
Zygote , Volume 17 , Issue 4 , November 2009 , pp. 315 - 320
Copyright
Copyright © Cambridge University Press 2009

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References

Anzur, M. & Buhr, M.M. (2006). Spontaneous uptake of exogenous DNA by bull spermatozoa. Theriogenology 65, 683–90.CrossRefGoogle Scholar
Atkinson, P.W., Hines, E.R., Beaton, S., Matthaei, K.I., Reed, K.C. & Bradley, M.P. (1991). Association of exogenous DNA with cattle and insect spermatozoa in vitro. Mol. Reprod. Dev. 29, 1–5.CrossRefGoogle ScholarPubMed
Bielfeld, P., Faredi, A., Zaneveld, L.J.D. & Dejomge, C.J. (1994). The zone pellucida-induced acrosome reaction of non-capacitated human spermatozoa. Fertil. Steril. 61, 536–41.CrossRefGoogle Scholar
Bradley, M.P., Geelan, A., Leitch, V. & Goldberg, E. (1996). Cloning, sequencing and characterization of LDH-C4 from a fox testis cDNA library. Mol. Reprod. Dev. 44, 452–9.3.0.CO;2-K>CrossRefGoogle ScholarPubMed
Bunch, D.O., Welch, J.E., Magyar, P.L., Eddy, E.M. & O'Brien, D.A. (1998). Glyceraldehyde 3-phosphate dehydrogenase-S protein distribution during mouse spermatogenesis. Biol. Reprod. 58, 834–41.CrossRefGoogle ScholarPubMed
Camaioni, A., Russo, M.A., Odorisio, T., Gandolfi, F., Fazio, V.M. & Siracusa, G. (1992). Uptake of exogenous DNA by mammalian spermatozoa specific localization of DNA on sperm heads. J. Reprod. Fertil. 96, 203–12.CrossRefGoogle ScholarPubMed
Castro, F.O., Hernandez, O., Uliver, C., Solano, R., Milanés, C., Aguilar, A., Pérez, A., De Armas, R., Herrwera, L. & De La Fuente, J. (1990). Introduction of foreign DNA into the spermatozoa of farm animals. Theriogenology 34, 1099–110.CrossRefGoogle Scholar
Celebi, C., Guillaudeux, T., Auvray, P., Vallet-Erdtmann, V. & Jégou, B. (2003). The making of “transgenic spermatozoa.” Biol. Reprod. 69, 1477–83.CrossRefGoogle Scholar
Celeghini, E.C.C., Arruda, R.P., Andrade, A.F.C., Nascimento, J. & Raphael, C.F. (2007). Practical techniques for bovine sperm simultaneous fluorimetric assessment of plasma, acrosomal and mitochondrial membranes. Reprod. Domest. Anim. 42, 479–88.CrossRefGoogle ScholarPubMed
Francolini, M., Lavitrano, M., Lamia, C.L., French, D., Frati, L., Cotelli, F. & Spadafora, C. (1993). Evidence for nuclear internalization of exogenous DNA into mammalian sperm cells. Mol. Reprod. Dev. 34, 133–9.CrossRefGoogle ScholarPubMed
Holt, C., Holt, W.V., Moore, H.D., Reed, H.B. & Curnock, R.M. (1997). Objectively measured boar sperm motility parameters correlate with the outcomes of on-farm inseminations: results of two fertility trials. J. Androl. 18, 312–23.CrossRefGoogle ScholarPubMed
Horan, R., Powell, R., Mcquaid, S., Gannon, F. & Houghton, J.A. (1991). Association of foreign DNA with porcine spermatozoa. Arch. Androl. 26, 83–92.CrossRefGoogle ScholarPubMed
Kjaestad, H., Ropstad, E. & Berg, K.A. (1993). Evaluation of spermatological parameters used to predict the fertility of frozen bull semen. Acta Vet. Scand. 34, 299–303.CrossRefGoogle ScholarPubMed
Lavitrano, M., Camaioni, A., Fazio, V.M., Dolci, S., Farace, M.G. & Spadafora, C. (1989). Sperm cells as vectors for introducing foreign DNA into eggs – genetic transformation of mice. Cell 57, 717–23.CrossRefGoogle ScholarPubMed
Maione, B., Pittogi, C., Achene, L., Lorenzini, R. & Spadafora, C. (1997) Activation of endonucleases in mature sperm cells upon interaction with exogenous DNA. DNA Cell. Biol. 16, 1087–97.CrossRefGoogle ScholarPubMed
Mori, C., Nakamura, N., Welch, J.E., Gotoh, H., Goulding, E.H., Fujioka, M. & Eddy, E.M. (1998). Mouse spermatogenic cell-specific type 1 hexokinase (mHk1-s) transcripts are expressed by alternative splicing from the mHk1 gene and the HK1-S protein is localized mainly in the sperm tail. Mol. Reprod. Dev. 49, 374–85.3.0.CO;2-K>CrossRefGoogle ScholarPubMed
Parrish, J.J., Susko-Parrish, J., Winer, M.A. & First, N.L. (1988). Capacitation of bovine sperm by heparin. Biol. Reprod. 38, 1171–80.CrossRefGoogle ScholarPubMed
Silva, P.N.F. & Gadella, B.M. (2006). Detection of damage in mammalian sperm cells. Theriogenology 65, 958–78.CrossRefGoogle ScholarPubMed
Smith, K. & Spadafora, C. (2005). Sperm-mediated gene transfer: applications and implications. Mol. Reprod. Dev. 25, 551–62.Google Scholar
Tanghe, S. Van Soon, A., Sterckx, V., Maes, D. & Kruif, A. (2002). Assessment of different sperm quality parameters to predict in vitro fertility of bulls. Reprod. Domest. Anim. 37, 127–32.CrossRefGoogle ScholarPubMed
Travis, A.J., Foster, J.A., Rosenbaum, N.A., Visconti, P.E., Gerton, G.L., Kopf, G.S. & Moss, S.B. (1998). Targeting of a germ cell-specific type 1 hexokinase lacking a porin-binding domain to the mitochondria as well as to the head and fibrous sheath of murine spermatozoa. Mol. Biol. Cell. 9, 263–76.CrossRefGoogle Scholar
Westhoff, D. & Kamp, G. (1997) Glyceraldehyde-3-phosphate dehydrogenase is bound to the fibrous sheath of mammalian spermatozoa. J. Cell Sci. 110, 1821–9.CrossRefGoogle Scholar