Recording and sorting live human sperm undergoing acrosome reaction

ZOPPINO FELIPE CAZLOS MARTIN; HALÓN NARCISO D; BUSTOS MATÍAS A; PAVAROTTI MARTÍN A; MAYORGA LUIS S.

FERTILITY AND STERILITY

ELSEVIER SCIENCE INC

Lugar: New York; Año: 2012

0015-0282

ABSTRACTOBJECTIVE: To develop and evaluate a method to detect acrosome reaction in live humansperm.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 fluoresceinisothiocyanate rapidly permeates into the acrosome when fusion pores open and stabilizes theacrosomal matrix, preventing the dispersal of the granule contents.CONCLUSIONS: Fluorescent Pisum sativum agglutinin can be used to visualize acrosomereaction in real time, to determine the percentage of sperm undergoing exocytosis uponstimulation, and to separate the population of reacting sperm by flow cytometry.