PMID- 20113445
OWN - NLM
STAT- MEDLINE
DCOM- 20110602
LR  - 20101122
IS  - 1439-0531 (Electronic)
IS  - 0936-6768 (Linking)
VI  - 45
IP  - 6
DP  - 2010 Dec
TI  - Comparison of the TBARS assay and BODIPY C11 probes for assessing lipid 
      peroxidation in red deer spermatozoa.
PG  - e360-8
LID - 10.1111/j.1439-0531.2009.01578.x [doi]
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 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.
CI  - (c) 2010 Blackwell Verlag GmbH.
FAU - Dominguez-Rebolledo, A E
AU  - Dominguez-Rebolledo AE
AD  - Biology of Reproduction Group, National Wildlife Research Institute (IREC) 
      (UCLM-CSIC-JCCM), Albacete, Spain.
FAU - Martinez-Pastor, F
AU  - Martinez-Pastor F
FAU - Fernandez-Santos, M R
AU  - Fernandez-Santos MR
FAU - del Olmo, E
AU  - del Olmo E
FAU - Bisbal, A
AU  - Bisbal A
FAU - Ros-Santaella, J L
AU  - Ros-Santaella JL
FAU - Garde, J J
AU  - Garde JJ
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Germany
TA  - Reprod Domest Anim
JT  - Reproduction in domestic animals = Zuchthygiene
JID - 9015668
RN  - 0 (4,4-difluoro-4-bora-3a,4a-diaza-s-indacene)
RN  - 0 (Boron Compounds)
RN  - 0 (Thiobarbituric Acid Reactive Substances)
SB  - IM
MH  - Animals
MH  - Boron Compounds/*chemistry
MH  - Cryopreservation/veterinary
MH  - *Deer
MH  - Lipid Peroxidation/*physiology
MH  - Male
MH  - Oxidative Stress
MH  - Reproducibility of Results
MH  - Semen Preservation/methods/veterinary
MH  - Sensitivity and Specificity
MH  - Spermatozoa/*physiology
MH  - Thiobarbituric Acid Reactive Substances/*metabolism
EDAT- 2010/02/02 06:00
MHDA- 2011/06/03 06:00
CRDT- 2010/02/02 06:00
PHST- 2010/02/02 06:00 [entrez]
PHST- 2010/02/02 06:00 [pubmed]
PHST- 2011/06/03 06:00 [medline]
AID - RDA1578 [pii]
AID - 10.1111/j.1439-0531.2009.01578.x [doi]
PST - ppublish
SO  - Reprod Domest Anim. 2010 Dec;45(6):e360-8. doi: 10.1111/j.1439-0531.2009.01578.x.