RT info:eu-repo/semantics/article
T1 Comparison of the TBARS Assay and BODIPY C11 Probes for Assessing Lipid Peroxidation in Red Deer Spermatozoa
A1 Domínguez Rebolledo, Álvaro Efrén
A1 Martínez Pastor, Felipe
A1 Fernández Santos, María Rocío
A1 Olmo de Medina, Enrique del
A1 Bisbal Vigo, Alfonso
A1 Ros Santaella, José Luis
A1 Garde López-Brea, Julián
A2 Biologia Celular
K1 Veterinaria
K1 Peroxidación lipídica
K1 Esperma
K1 Ciervo rojo
AB Several methods are used to measure lipid peroxidation (LPO) in spermatozoa. The objective of this study was comparing the thiobarbituric acid reactive species (TBARS) method and the BODIPY 581/591 C11 (B581) and BODIPY 665/676 C11 (B665) fluorescent probes to measure induced peroxidative damage in thawed epididymal spermatozoa from Iberian red deer. Samples from three males were thawed, pooled, diluted in PBS, incubated at room temperature and assessed at 0, 3, 6 and 24 h under different experimental conditions: Control, hydrogen peroxide (H2O2) 0.1 mm or 1 mm, or tert‐butyl hydroperoxide (TBH) 0.1 mm or 1 mm. LPO was assessed by the TBARS assay [malondialdehyde (MDA) detection] and by the fluorescence probes B581 and B665 (microplate fluorimeter and flow cytometry). Increasing MDA levels were only detectable at 1 mm of TBH or H2O2. Both fluorescence probes, measured with fluorometer, detected significant increases of LPO with time in all treatments, except Control. Flow cytometry allowed for higher sensitivity, with both probes showing a significant linear relationship of increasing LPO with time for all oxidizing treatments (p < 0.001). All methods showed a good agreement, except TBARS, and flow cytometry showed the highest repeatability. Our results show that both B581 and B665 might be used for LPO analysis in Iberian red deer epididymal spermatozoa, together with fluorometry or flow cytometry. Yet, the TBARS method offered comparatively limited sensitivity, and further research must determine the source of that limitation.
PB John Wiley & Sons
YR 2019
FD 2019-04-15
LK http://hdl.handle.net/10612/10225
UL http://hdl.handle.net/10612/10225
NO Reproduction in Domestic Animals, 2010, vol. 45, n. 6
NO P. 360-368
DS BULERIA. Repositorio Institucional de la Universidad de León
RD 26-abr-2024