Uso da lectina do quiabo (Abelmoschus esculentus) como marcador da célula espermática ovina

Abstract

The lectins are employeds as surface markers in the plasma membrane of spermatozoa due to its binding to saccharides thereof. The aim is to seek the connection location and the collaboration of the lectin quiado (Abelmoschus esculentus) in sperm sheep. The lectin A. esculentus was purified as described in Smith et al. (2012). 8 ejaculates of ovines were used. The samples were diluted in Tris-egg yolk pattern plus diluent different concentrations of lectins according to treatment: T1 -0μg lectin; T2 1μg lectin; T3 -3μg lectin; T4 -5μg lectin. The samples were stored at 5°C. For reviews on sperm periods 0h, 2h and 24 hours they were heated to 37°C. For the analysis of membrane integrity, mitochondria, fluidity and ROS, we used flow cytometry. To evaluate the kinetics of sperm cells, we used the Computer-Assisted Sperm Analysis system (CASA). The marking of the lectin in the sperm cell was made with FITC and observed by Confocal Microscope Inverted Scanning Laser and observed that the fluorescence was detected in the intermediate part of the spermatozoon. In no conditioning period difference was observed in relation to membrane integrity, mitochondria and sperm flow. At 0 hour period in sperm kinetic displacement and the speed they were lower T4 than in T3 (P <0.05). In the assessment of 2 hours T2 was higher than others in the displacement and sperm velocity (P <0.05). After conditioning for 24 h at 5°C sperm kinetics was higher in T3 compared to T4 not differing from the other treatments. This finding allows that seminal assessment can be made up to 24 hours without loss in the seminal characteristics when added up to 3μg lectin to 5°C. In addition to evidence that its binding site is in the middle piece of sperm ovines.