34 resultados para CAPACITATION


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Because the routine use of frozen semen has some limitation that don´t permit its use in a large-scale, it is necessary to use the cooled semen. The equine cooled semen is normally used to enable that a genetic material with high quality be spread over long distances. When it reaches the temperature of refrigeration, the sperm metabolic activity decreases and the free radicals formation minimize. These ones cause irreversible damages to the sperm cells and, so, its lower formation is very advantageous. However, when we manipulate the semen using conservation techniques, like refrigeration, it is necessary to be aware about the sperm characteristics and fragilities, because, if performed erroneously, this technique can be harmful to the sperm function as well as to the time of sperm capacitation and acrosome reaction. It is necessary that cooling rate is slow and that the time and the storage temperature of the sperm obey the ranges that are already established. Moreover, we should make use of diluents and obtain the ideal sperm dilution, so that its use can be optimized. It´s also important to emphasize that to obtain good fertility rates, the semen, after processed (collected and diluted) must be conditioned in recipients specially developed for this purpose

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Methods of semen cryopreservation allow changes in spermatic cells, such as damage in plasma and acrossomal membrane and modifications in mitochondrial function due to a disorder in the lipidic bilayer. For effective oocyte fertilization, spermatozoa require functional competent membranes, and intact organelles, acrosome and DNA. However, most laboratory methods used to evaluate semen quality are not highly correlated with fertilizing capacity. The discovery of a variety of fluorochromes and compounds conjugated to fluorescent probes has enabled an accurate assessment of the viability, integrity and function of spermatozoa. Among the most used probes that label the various compartments of the sperm cell there are the membrane impermeable fluorescent dyes to test the membrane integrity, as well as acylated dyes that pass the intact membrane. For the acrossomal integrity the most commonly used method is lectins labeled by a fluorescent probe. The acrosome reaction and spermatic capacitation is detected by the evaluation of membrane architecture and disorder of lipids in plasma membrane. Mitochondrial function can be determined using markers for their aerobic activity. The DNA status of spermatozoa has been determined using the metachromatic properties of Acridine Orange, and the DNA fragmentation can also be assessed by TUNEL assay. Finally, DNA condensation is analyzed using a single cell DNA gel electrophoresis assay that indicates DNA compactation. This monograph aims to compile the various tests used to detect damaged spermatozoa under cryopreservation methods, searching for improve the predictive value of semen analysis with the intention of a successful conception

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)