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Real-Time Imaging of Acrosomal Calcium Dynamics and Exocytosis in Live Mouse Sperm
Published on: October 13, 2023
Recording and sorting live human sperm undergoing acrosome reaction.
Felipe Carlos Martín Zoppino1, Narciso D Halón, Matías A Bustos
1Instituto de Histología y Embriología Dr Mario H Burgos, Consejo Nacional de Investigaciones Científicas y Técnicas - Universidad Nacional de Cuyo, School of Medicine, National University of Cuyo, Mendoza, Argentina.
Fertility and Sterility
|April 13, 2012
Summary
A new method uses fluorescent Pisum sativum agglutinin to detect the acrosome reaction in live human sperm, enabling real-time visualization and separation of reacting sperm populations.
Area of Science:
- Reproductive Biology
- Sperm Physiology
Background:
- Detecting acrosome reaction (AR) in live human sperm is crucial for fertility assessments.
- Current methods for evaluating AR present challenges in real-time analysis.
Purpose of the Study:
- To develop and validate a novel method for detecting the acrosome reaction in live human sperm.
- To enable real-time visualization and quantification of sperm undergoing acrosome reaction.
Main Methods:
- Prospective study design utilizing human semen samples from healthy donors.
- Acrosome reaction assays were performed with fluorescence assessment.
- Pisum sativum agglutinin conjugated to fluorescein isothiocyanate was used for detection.
Main Results:
- Fluorescent Pisum sativum agglutinin rapidly permeates the acrosome upon fusion pore opening during AR.
- The fluorescent marker stabilizes the acrosomal matrix, preventing content dispersal.
- This allows for the evaluation of acrosomal exocytosis in live sperm.
Conclusions:
- Fluorescent Pisum sativum agglutinin provides a reliable method for real-time visualization of AR.
- The technique allows for accurate determination of the percentage of sperm undergoing exocytosis.
- Reacting sperm populations can be effectively separated using flow cytometry.
