Flow-sorted ram spermatozoa are highly susceptible to hydrogen peroxide damage but are protected by seminal plasma and catalase

Leahy, T., Celi, P., Bathgate, R., Evans, G., Maxwell, W. M. C. and Marti, J. I. (2010) Flow-sorted ram spermatozoa are highly susceptible to hydrogen peroxide damage but are protected by seminal plasma and catalase. Reproduction Fertility and Development, 22 7: 1131-1140. doi:10.1071/RD09286


Author Leahy, T.
Celi, P.
Bathgate, R.
Evans, G.
Maxwell, W. M. C.
Marti, J. I.
Title Flow-sorted ram spermatozoa are highly susceptible to hydrogen peroxide damage but are protected by seminal plasma and catalase
Journal name Reproduction Fertility and Development   Check publisher's open access policy
ISSN 1031-3613
1448-5990
Publication date 2010
Sub-type Article (original research)
DOI 10.1071/RD09286
Volume 22
Issue 7
Start page 1131
End page 1140
Total pages 10
Editor Tony Flint
Place of publication Canberra, ACT, Australia
Publisher CSIRO Publishing
Collection year 2011
Language eng
Formatted abstract
To determine whether flow sorting increased the susceptibility of spermatozoa to reactive oxygen species (ROS), ram semen was either diluted with Tris medium (100 × 106 spermatozoa mL−1; D) or highly diluted (106 spermatozoa mL−1) before being centrifuged (DC) at 750g for 7.5 min at 21°C or flow-sorted (S) before cryopreservation. Thawed spermatozoa were resuspended in graded concentrations of hydrogen peroxide to induce oxidative stress. In Experiment 1, following exposure to 30 or 45 μM hydrogen peroxide (H2O2), the total motility (%) of DC (41.0 ± 7.3 or 25.7 ± 6.7, respectively) and S spermatozoa (33.8 ± 6.3 or 20.1 ± 6.3, respectively) was lower (P < 0.001) than that of D spermatozoa (58.7 ± 5.6 or 44.5 ± 6.7, respectively). In Experiment 2, supplementation of samples containing H2O2 with catalase (150 IU mL−1) or seminal plasma proteins (4 mg protein per 108 spermatozoa) negated oxidative stress, resulting in comparable values to samples receiving no H2O2in terms of the proportion of spermatozoa with stable plasmalemma (as determined using merocyanine-540 and Yo-Pro-1) in the D and S groups, the proportion of viable, acrosome-intact spermatozoa (as determined by fluorescein isothiocyanate and propidium iodide staining) in the D group and the motility of control (undiluted) and S spermatozoa. Neither H2O2 nor sperm type (i.e. D, DC or S) had any effect on intracellular concentrations of ROS. These results show that flow sorting increases the susceptibility of spermatozoa to ROS, but the inclusion of anti-oxidants or seminal plasma as part of the sorting protocol improves resistance to oxidative stress.
© CSIRO 2010
Keyword Cryopreservation
Flow cytometry
Reactive oxygen species
Sex sorting
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Non HERDC
School of Veterinary Science Publications
 
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Created: Mon, 04 Apr 2011, 14:04:55 EST by Dr Tamara Leahy on behalf of School of Veterinary Science