2024-03-28T08:52:09Zhttp://buleria.unileon.es/oai/requestoai:buleria.unileon.es:10612/102252020-12-10T09:01:37Zcom_10612_17col_10612_18
BULERIA. Repositorio Institucional de la Universidad de León
author
Domínguez Rebolledo, Álvaro Efrén
author
Martínez Pastor, Felipe
author
Fernández Santos, María Rocío
author
Olmo de Medina, Enrique del
author
Bisbal Vigo, Alfonso
author
Ros Santaella, José Luis
author
Garde López-Brea, Julián
other
Biologia Celular
2019-04-14T23:26:11Z
2019-04-14T23:26:11Z
2019-04-15
Reproduction in Domestic Animals, 2010, vol. 45, n. 6
https://onlinelibrary.wiley.com/doi/full/10.1111/j.1439-0531.2009.01578.x
http://hdl.handle.net/10612/10225
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.
Veterinaria
Comparison of the TBARS Assay and BODIPY C11 Probes for Assessing Lipid Peroxidation in Red Deer Spermatozoa
info:eu-repo/semantics/article
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URL
https://buleria.unileon.es/bitstream/10612/10225/1/37%20Comparison%20of%20the%20TBARS%20Assay%20and%20BODIPY%20C11%20ReprodDomestAnim_045_e0360_2010.pdf
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37 Comparison of the TBARS Assay and BODIPY C11 ReprodDomestAnim_045_e0360_2010.pdf
URL
https://buleria.unileon.es/bitstream/10612/10225/3/37%20Comparison%20of%20the%20TBARS%20Assay%20and%20BODIPY%20C11%20ReprodDomestAnim_045_e0360_2010.pdf.txt
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37 Comparison of the TBARS Assay and BODIPY C11 ReprodDomestAnim_045_e0360_2010.pdf.txt