Bovine sperm cells viability during incubation with or without exogenous DNA.
SummaryThe 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 x 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.
Feitosa
Weber Beringui
WB
Department of Animal Reproduction, College of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil.
Milazzotto
Marcella Pecora
MP
Simões
Renata
R
Rovegno
Mariana
M
Nicacio
Alessandra Coralo
AC
Nascimento
Aníbal Ballarotti
AB
Gonçalves
José Sérgio Arruda
JS
Visintin
José Antonio
JA
Assumpção
Mayra Elena Ortiz D'Avila
ME
D000177:Acrosome / Q000378:metabolism; D000818:Animals; D002417:Cattle / Q000378:metabolism*; D002470:Cell Survival; D004247:DNA / Q000378:metabolism*; D018014:Gene Transfer Techniques; D008297:Male; D053078:Membrane Potential, Mitochondrial; D013094:Spermatozoa / Q000166:cytology*
D004247:DNA