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Bovine sperm cells viability during incubation with or without exogenous DNA

  • Weber Beringui Feitosa (a1), Marcella Pecora Milazzotto (a1) (a2), Renata Simões (a1), Mariana Rovegno (a1), Alessandra Coralo Nicacio (a1) (a3), Aníbal Ballarotti Nascimento (a1) (a4), José Sérgio Arruda Gonçalves (a1), José Antonio Visintin (a1) and Mayra Elena Ortiz D'Ávila Assumpção (a5) (a1)...

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.

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Corresponding author

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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Bovine sperm cells viability during incubation with or without exogenous DNA

  • Weber Beringui Feitosa (a1), Marcella Pecora Milazzotto (a1) (a2), Renata Simões (a1), Mariana Rovegno (a1), Alessandra Coralo Nicacio (a1) (a3), Aníbal Ballarotti Nascimento (a1) (a4), José Sérgio Arruda Gonçalves (a1), José Antonio Visintin (a1) and Mayra Elena Ortiz D'Ávila Assumpção (a5) (a1)...

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