| Comparison of the TBARS assay and BODIPY C11 probes for assessing lipid peroxidation in red deer spermatozoa. | |
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MedLine Citation:
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PMID: 20113445 Owner: NLM Status: In-Process |
Abstract/OtherAbstract:
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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 C(11) (B581) and BODIPY 665/676 C(11) (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 (H(2)O(2) ) 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 H(2)O(2). 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. |
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Authors:
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Á E Domínguez-Rebolledo; F Martínez-Pastor; M R Fernández-Santos; E del Olmo; A Bisbal; J L Ros-Santaella; J J Garde |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Reproduction in domestic animals = Zuchthygiene Volume: 45 ISSN: 1439-0531 ISO Abbreviation: Reprod. Domest. Anim. Publication Date: 2010 Dec |
Date Detail:
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Created Date: 2010-11-22 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9015668 Medline TA: Reprod Domest Anim Country: Germany |
Other Details:
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Languages: eng Pagination: e360-8 Citation Subset: IM |
Copyright Information:
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© 2010 Blackwell Verlag GmbH. |
Affiliation:
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Biology of Reproduction Group, National Wildlife Research Institute (IREC) (UCLM-CSIC-JCCM), Albacete, Spain. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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