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May 15, 2007Journal of Animal Science

Use of fluorescence-activated flow cytometry to determine membrane lipid peroxidation during hypothermic liquid storage and freeze-thawing of viable boar sperm loaded with 4, 4-difluoro-5-(4-phenyl-1,3-butadienyl)-4-bora-3a,4a-diaza-s-indacene-3-undecanoic acid1

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Authors

HGH. D. GuthrieAgricultural Research Service GWG. Rickey WelchAgricultural Research Service

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Implication

Experimental study reveals low basal lipid peroxidation in stored and frozen boar sperm, indicating that cellular decay arises primarily from iron-catalyzed oxidative stress.

Key Points

To evaluate membrane lipid peroxidation in viable boar spermatozoa during hypothermic liquid storage and freeze-thawing using the lipophilic fluorescent probe C11-BODIPY(581/591) and flow cytometry.
Loaded Percoll-washed, propidium iodide-negative (viable) boar spermatozoa with C11-BODIPY(581/591) and tested oxidation specificity against reactive oxygen species generators (FeSO4 plus sodium ascorbate [FeAc], menadione, H2O2) and metal chelators (EDTA, deferoxamine mesylate).
Quantified basal and FeAc-induced BODIPY oxidation using flow cytometry after liquid storage at 17°C (day 1 and day 5) and post cryopreservation freeze-thawing.
FeAc incubation increased the proportion of sperm containing oxidized BODIPY to >70% (P < 0.01) and raised fluorescence intensity 5- to 10-fold (P < 0.05), which depressed motility (P < 0.05) without altering viability.
Metal chelators EDTA and deferoxamine mesylate at 3 and 9 µM inhibited FeAc-induced BODIPY oxidation and preserved sperm motility.
Basal spontaneous oxidation was low in both liquid-stored (1.3%) and thawed viable sperm (3.4%), though viable cells remained susceptible to FeAc-induced lipid peroxidation.

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Guthrie et al. (2007) studied this question.

synapsesocial.com/papers/6aa38940fd19ae9969b146be https://doi.org/10.2527/jas.2006-787

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Implication

Experimental study reveals low basal lipid peroxidation in stored and frozen boar sperm, indicating that cellular decay arises primarily from iron-catalyzed oxidative stress.

Use of fluorescence-activated flow cytometry to determine membrane lipid peroxidation during hypothermic liquid storage and freeze-thawing of viable boar sperm loaded with 4, 4-difluoro-5-(4-phenyl-1,3-butadienyl)-4-bora-3a,4a-diaza-s-indacene-3-undecanoic acid1

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Cite This Study

Use of fluorescence-activated flow cytometry to determine membrane lipid peroxidation during hypothermic liquid storage and freeze-thawing of viable boar sperm loaded with 4, 4-difluoro-5-(4-phenyl-1,3-butadienyl)-4-bora-3a,4a-diaza-s-indacene-3-undecanoic acid1

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