IMPACT OF COPPER TO THE CHELATION EFF ECT OF BOVINE SERUM ALBUMIN AND SPERMATOZOA MOTILITY PARAMETER IN VITRO

The target of this in vitro study was to analyse the influence of copper on the spermatozoa motility in the presence of bovine serum albumin (BSA) as culture medium and to provide additional information on the interaction between serum albumin and copper (II) chloride (CuCl₂). The spermatozoa motility parameters was determined after exposure of CuCl₂ (3.9; 7.8; 15.6; 31.2; 62.5; 125; 250; 500; 1000 μmol/L) using the Sperm Vision^TM CASA (Computer Assisted Semen Analyzer) system during different time periods (Time 0 h, 1 h, 2 h and 24). The culture medium containing 20% BSA, triladyl and 5% glucose increased the overall percentage of spermatozoa motility after 1 h of cultivation. The percentage of motility spermatozoa significantly (P<0.001) decreased after 2 h of cultivation at the concentrations ≥ 250 μmol/L of CuCl₂ in comparison with the control group (without CuCl₂ administration). The experimental administration at the doses ≤ 31.2 μmol/L of CuCl₂ stimulated the overall of motile spermatozoa (Time 24 h). Identical
spermatozoa motility was detected also for the percentage of progressive motile spermatozoa during all time periods. Parameter of distance average path (DAP) showed increase in all CuCl₂ addition groups in comparison with the control group at Time 1 h. Concentration 125 μmol/L of CuCl₂ in various time periods of cultivation act stimulating on spermatozoa motility, but later (Time
24 h) inhibitory. Evaluation of velocity average path, showed similar results as for DAP. Measurement of the amplitude of lateral displacement (ALH) at Time 0 h as well as at Time 1 h was higher in all the experimental groups compared to the control group, but the differences were not significant (P>0.05). The experimental administration at the doses ≤ 62.5 μmol/L of CuCl₂ stimulated
ALH during 24 h of cultivation. The results suggest that adding energy and protein substrate to the culture medium increases the spermatozoa motility parameters also the presence of high doses (≥ 125 μmol/L) of copper ions during short-term periods (Time 0 h, 1 h). Concurrently BSA maintained motility of spermatozoa (≤ 31.20 μmol/L of CuCl₂) during the long-term (Time 24 h) of cultivation, which confirms the protective effect of albumin binding to the copper ions.