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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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