Recording and sorting live human sperm undergoing acrosome reaction

ZOPPINO FCM, HALÓN ND, BUSTOS MA, PAVAROTTI MA Y MAYORGA LS.

FERTILITY AND STERILITY

ELSEVIER SCIENCE INC

Año: 2012

0015-0282

OBJECTIVE: To develop and evaluate a method to detect acrosome reaction in live human sperm. DESIGN: Prospective study. SETTING: Basic research laboratory. PATIENT(s): Human semen samples with normal parameters obtained from healthy donors. INTERVENTIONS: Acrosome reaction assays. MAIN OUTOME: Fluorescence assessment of acrosome reaction. RESULTS: Evaluating acrosomal exocytosis in live human sperm is challenging. In this study, we report that in reacting sperm, Pisum sativum agglutinin conjugated to fluorescein isothiocyanate rapidly permeates into the acrosome when fusion pores open and stabilizes the acrosomal matrix, preventing the dispersal of the granule contents. CONCLUSIONS: Fluorescent Pisum sativum agglutinin can be used to visualize acrosome reaction in real time, to determine the percentage of sperm undergoing exocytosis upon stimulation, and to separate the population of reacting sperm by flow cytometry. reaction in real time, to determine the percentage of sperm undergoing exocytosis upon stimulation, and to separate the population of reacting sperm by flow cytometry. isothiocyanate rapidly permeates into the acrosome when fusion pores open and stabilizes the acrosomal matrix, preventing the dispersal of the granule contents. CONCLUSIONS: Fluorescent Pisum sativum agglutinin can be used to visualize acrosome reaction in real time, to determine the percentage of sperm undergoing exocytosis upon stimulation, and to separate the population of reacting sperm by flow cytometry. reaction in real time, to determine the percentage of sperm undergoing exocytosis upon stimulation, and to separate the population of reacting sperm by flow cytometry. Pisum sativum agglutinin conjugated to fluorescein isothiocyanate rapidly permeates into the acrosome when fusion pores open and stabilizes the acrosomal matrix, preventing the dispersal of the granule contents. CONCLUSIONS: Fluorescent Pisum sativum agglutinin can be used to visualize acrosome reaction in real time, to determine the percentage of sperm undergoing exocytosis upon stimulation, and to separate the population of reacting sperm by flow cytometry. reaction in real time, to determine the percentage of sperm undergoing exocytosis upon stimulation, and to separate the population of reacting sperm by flow cytometry. Pisum sativum agglutinin can be used to visualize acrosome reaction in real time, to determine the percentage of sperm undergoing exocytosis upon stimulation, and to separate the population of reacting sperm by flow cytometry.