OBJECTIVE: To develop and evaluate a method to detect acrosome reaction (AR) in live human sperm. DESIGN: Prospective study. SETTING: Basic research laboratory. PATIENT(S): Human semen samples with normal parameters obtained from healthy donors. INTERVENTION(S): Acrosome reaction assays. MAIN OUTCOME MEASURE(S): Fluorescence assessment of AR. RESULT(S): 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. CONCLUSION(S): Fluorescent Pisum sativum agglutinin can be used to visualize AR in real time, to determine the percentage of sperm undergoing exocytosis upon stimulation, and to separate the population of reacting sperm by flow cytometry.
OBJECTIVE: To develop and evaluate a method to detect acrosome reaction (AR) in live human sperm. DESIGN: Prospective study. SETTING: Basic research laboratory. PATIENT(S): Human semen samples with normal parameters obtained from healthy donors. INTERVENTION(S): Acrosome reaction assays. MAIN OUTCOME MEASURE(S): Fluorescence assessment of AR. RESULT(S): 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. CONCLUSION(S): Fluorescent Pisum sativum agglutinin can be used to visualize AR in real time, to determine the percentage of sperm undergoing exocytosis upon stimulation, and to separate the population of reacting sperm by flow cytometry.
Authors: Guillermina M Luque; Tomas Dalotto-Moreno; David Martín-Hidalgo; Carla Ritagliati; Lis C Puga Molina; Ana Romarowski; Paula A Balestrini; Liza J Schiavi-Ehrenhaus; Nicolas Gilio; Dario Krapf; Pablo E Visconti; Mariano G Buffone Journal: J Cell Physiol Date: 2018-06-28 Impact factor: 6.384
Authors: María F Quevedo; Ornella Lucchesi; Matías A Bustos; Cristian A Pocognoni; Paola X De la Iglesia; Claudia N Tomes Journal: J Biol Chem Date: 2016-09-09 Impact factor: 5.157
Authors: Ruiming Zhao; Kelleigh Kennedy; Gerardo A De Blas; Gerardo Orta; Martín A Pavarotti; Rodolfo J Arias; José Luis de la Vega-Beltrán; Qufei Li; Hui Dai; Eduardo Perozo; Luis S Mayorga; Alberto Darszon; Steve A N Goldstein Journal: Proc Natl Acad Sci U S A Date: 2018-11-26 Impact factor: 11.205
Authors: Leonardo E Pelletán; Laila Suhaiman; Cintia C Vaquer; Matías A Bustos; Gerardo A De Blas; Nicolas Vitale; Luis S Mayorga; Silvia A Belmonte Journal: J Biol Chem Date: 2015-02-20 Impact factor: 5.157