Semen quality evaluation and determination of its fertilizing ability are essential aspects in the practice of assisted reproduction in domestic species. Fertilization is a multifunctional process, thus several tests related to sperm functional competence and multiparametric data analysis are required for a proper evaluation. Despite many studies are available in the literature and much progress has been made, at present a set of tests suitable for the prediction of fertilizing ability is still not available. Flow cytometry (FCM), allowing the analysis of different parameters on several sperm cells in short time, can provide a fair assessment of semen potential fertility. In this study different quality parameters of bull frozen-thawed semen were evaluated by FCM and by computer assisted sperm analysis (CASA). Frozen semen from 18 Holstein-Friesian bulls, 10 batches each, was assessed. The following parameters were tested by FCM: sperm viability (SYBR-14/propidium iodide (PI) staining), acrosomal status (FITC-PNA/PI staining), membrane lipid disorder (Merocyanine 540 hydrophobic dye), lipid peroxidation (BODIPY (581/591) C11 lipophilic probe), mitochondrial membrane potential (JC-1 potentiometric probe). Sperm kinetic was assessed by CASA. The mean values of measured parameters, standard deviations and minimum and maximum values were: proportion of viable sperm: 41.73±10.57% (16.26 to 66.44); live acrosomeintact sperm: 34.34±10.52% (8.52 to 56.55); live low lipid disorder sperm: 22.54±7.62% (6.97 to 49.84); live peroxidized sperm: 0.55±0.92% (0.00 to 8.49); live peroxidized sperm after promoter (FeSO4) treatment: 4.28±2.93% (0.76 to 16.41); high mitopotential sperm: 43.89±10.74% (9.78 to 67.09); motile sperm: 63.43±14.32% (14.50 to 88.50). Values were in general fairly consistent with levels found in the literature. Sperm viability was significantly correlated (P<0.0001) with low membrane lipid disorder (r=0.68) and acrosome-intactness (r=0.62), both in live sperm, and with motility (r=0.59). Moreover high mitopotential showed moderate but highly significant (P<0.0001) correlations with low lipid disorder in live sperm (r=0.51) and with motility (r=0.40). FCM can be considered a helpful tool for quick multiparametric evaluation of frozen-thawed bull semen. At present the estimation of the correlations between in vivo fertility and FCM semen quality parameters is in progress.
Flow cytometric evaluation of functional parameters in bovine semen.
Pizzi F;Gliozzi TM
2013
Abstract
Semen quality evaluation and determination of its fertilizing ability are essential aspects in the practice of assisted reproduction in domestic species. Fertilization is a multifunctional process, thus several tests related to sperm functional competence and multiparametric data analysis are required for a proper evaluation. Despite many studies are available in the literature and much progress has been made, at present a set of tests suitable for the prediction of fertilizing ability is still not available. Flow cytometry (FCM), allowing the analysis of different parameters on several sperm cells in short time, can provide a fair assessment of semen potential fertility. In this study different quality parameters of bull frozen-thawed semen were evaluated by FCM and by computer assisted sperm analysis (CASA). Frozen semen from 18 Holstein-Friesian bulls, 10 batches each, was assessed. The following parameters were tested by FCM: sperm viability (SYBR-14/propidium iodide (PI) staining), acrosomal status (FITC-PNA/PI staining), membrane lipid disorder (Merocyanine 540 hydrophobic dye), lipid peroxidation (BODIPY (581/591) C11 lipophilic probe), mitochondrial membrane potential (JC-1 potentiometric probe). Sperm kinetic was assessed by CASA. The mean values of measured parameters, standard deviations and minimum and maximum values were: proportion of viable sperm: 41.73±10.57% (16.26 to 66.44); live acrosomeintact sperm: 34.34±10.52% (8.52 to 56.55); live low lipid disorder sperm: 22.54±7.62% (6.97 to 49.84); live peroxidized sperm: 0.55±0.92% (0.00 to 8.49); live peroxidized sperm after promoter (FeSO4) treatment: 4.28±2.93% (0.76 to 16.41); high mitopotential sperm: 43.89±10.74% (9.78 to 67.09); motile sperm: 63.43±14.32% (14.50 to 88.50). Values were in general fairly consistent with levels found in the literature. Sperm viability was significantly correlated (P<0.0001) with low membrane lipid disorder (r=0.68) and acrosome-intactness (r=0.62), both in live sperm, and with motility (r=0.59). Moreover high mitopotential showed moderate but highly significant (P<0.0001) correlations with low lipid disorder in live sperm (r=0.51) and with motility (r=0.40). FCM can be considered a helpful tool for quick multiparametric evaluation of frozen-thawed bull semen. At present the estimation of the correlations between in vivo fertility and FCM semen quality parameters is in progress.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
