Actividad proteolitica de la Acrosina como parametro para evaluar el estado Acrosomico y predecir el potencial de fertilidad del espermatozoide humano / Proteolitic activity of Acrosine as a parameter to evaluate the acrosomic states and predict fertility of human spermatozoid

El presente trabajo fue orientado a la identificacion de un metodo de extraccion de la Acrosina control de la densidad espermatica despues de la extraccion, control de calidad para la eliminacion de contaminantes y el estudio de parametros cineticos de la enzima (concentracion de substrato, enzima, pH, temperatura, tiempo de reaccion). Se estudiaron 104 muestras de liquido seminal, los cuales fueron clasificados de acuerdo a criterios de la OMS. En todos los grupos clasificados se determinaron la actividad catalitica de la Acrosina y los parametros seminologicos. Se concluye que la determinacion de la actividad catalitica de la Acrosina discrimina entre los acrosomas indemnes de los espermatozoides normales; y los acrosomas con alteraciones de las membranas acrosomicas en espermios defectuosos por lo tanto evalua la capacidad de fertilidad del espermatozoide humano, siendo un importante parametro adicional el simograma de rutina, y un requisito para la fertilizacion in vitro e inseminacion artificila.

Immunodetection of acrosin during the acrosome reaction of hamster, guinea-pig and human spermatozoa

Mammalian sperm acrosomes contain a trypsin-like protease called acrosin which causes limited and specific hydrolysis of the extracellular matrix of the mammalian egg, the zona pellucida. Acrosin was localized on hamster, guinea-pig and human sperm using monoclonal and polyclonal antibodies to human acrosin labelled with colloidal gold. This was visualized directly with transmission electron microscopy, and with light and scanning microscopy after silver enhancement of the colloidal gold probe. Four distinct labelling patterns were found during capacitation and the acrosome reaction in hamster and guinea-pig spermatozoa, and three patterns were found in human spermatozoa. In the hamster, acrosin was not detected on the inner acrosomal surface after the completion of the acrosome reaction, thus correlating with the observation that hamster spermatozoa lose the ability to penetrate the zona after the acrosome reaction. With guinea-pig and human spermatozoa, acrosin was still detected after the completion of the acrosome reaction, thus correlating with the observation that acrosome reacted guinea-pig spermatozoa bind to and penetrate the zona pellucida