Quantitative determination of superoxide dismutase in crioconserved donkey semen by using a commercial kit [Tuscany]
2006
Ducci, M. | Felicioli, A. | Sighieri, C. | Rota, A. | Martelli, F. | Gazzano, A. | Pacchini, S. | Ducci, C.
Superoxide dismutases (SODs) are metalloenzymes that catalyze the dismutation of the superoxide anion to oxygen peroxide and molecular oxygen by redox cycling involving a metal ion. Superoxide dismutase is an important enzyme in the antioxidant cellular pathway, so the amount of SOD in cell and in the extracellular environment has outcomes in prevention of aging and disease. The oxide anion dismutation catalyzed by SOD in a very fast reaction (2 x 10E9 ME-1secE-1 turnover) and one unit of SOD is defined as the amount of enzyme needed to exhibit 50% dismutation of the superoxide radical. Several kits are available for SOD activity evaluation in plasma, serum, erythrocyte and cell lysate. The aim of this investigation was to assay SOD activity in seminal plasma and spermatozoa of Amiata Donkey by a commercial kit. The Amiata Donkey is an endangered native breed and SOD evaluation could contribute to the development of new semen conservation and quality evaluation methods. SOD was detected in samples of both seminal plasma and semen. SOD in seminal plasma resulted to be 10 time more concentrate (235,65 IU/ml) than in spermatozoa (19,15 IU/ml). Concentration differences between the two samples could be a real sample intrinsic difference as well they could be due to the spermatozoa protein extraction method. Spermatozoa underwent to a cold-shock treatment so that the subsequent release into the medium of the acrosomial proteinase could have, as a consequence, the partial inactivation of SOD; as well, the same treatment could be not sufficient for the complete membrane disruption. The kit resulted suitable for the assay of SOD in the ejaculate of Amiata Donkey in the dilution range (1/110 - 1/140) in spermatozoa (with a concentration of 575 x 10E6 spermatozoa/ml) and (1/1400 - 1/1700) for the seminal plasma
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