J Tesarik1, C Mendoza, A Carreras. 1. Center for Reproductive Biology and Medicine, American Hospital of Paris, Neuilly sur Seine, France.
Abstract
OBJECTIVE: To test a new method for evaluation of stimulus-induced acrosome reaction (AR). To determine whether this method, based on the definition of a specific staining pattern of recently reacted spermatozoa, brings an advantage of increased sensitivity as compared with a standard procedure. DESIGN: The hypothesis that a specific staining pattern with fluorescein-labeled Pisum sativum agglutinin corresponds to recently acrosome-reacted spermatozoa was tested by analyzing the frequency of this pattern at different time points after addition of ionophore A23187, an artificial AR inducer. The AR results obtained with a new method based on this relationship were compared with those obtained with a standard method of AR evaluation. SETTING: Private hospital, public research center, and a university-based research laboratory. PARTICIPANTS: Healthy sperm donors. MAIN OUTCOME MEASURE: Changes in the frequency of specific sperm-staining patterns. RESULTS: A specific sperm-staining pattern with P. sativum agglutinin was shown to be associated with a recently occurred AR, whereas the absence of staining was typical of reactions having occurred a longer time ago. This phenomenon was used to define a formula for fast AR measure after stimulus addition. An increased sensitivity provided by this formula as compared with a standard evaluation was demonstrated. CONCLUSIONS: Fast AR measure is a simple, easy-to-perform method for AR evaluation. It is particularly suitable for testing the effects of rapidly acting stimuli.
OBJECTIVE: To test a new method for evaluation of stimulus-induced acrosome reaction (AR). To determine whether this method, based on the definition of a specific staining pattern of recently reacted spermatozoa, brings an advantage of increased sensitivity as compared with a standard procedure. DESIGN: The hypothesis that a specific staining pattern with fluorescein-labeled Pisum sativum agglutinin corresponds to recently acrosome-reacted spermatozoa was tested by analyzing the frequency of this pattern at different time points after addition of ionophore A23187, an artificial AR inducer. The AR results obtained with a new method based on this relationship were compared with those obtained with a standard method of AR evaluation. SETTING: Private hospital, public research center, and a university-based research laboratory. PARTICIPANTS: Healthy sperm donors. MAIN OUTCOME MEASURE: Changes in the frequency of specific sperm-staining patterns. RESULTS: A specific sperm-staining pattern with P. sativum agglutinin was shown to be associated with a recently occurred AR, whereas the absence of staining was typical of reactions having occurred a longer time ago. This phenomenon was used to define a formula for fast AR measure after stimulus addition. An increased sensitivity provided by this formula as compared with a standard evaluation was demonstrated. CONCLUSIONS: Fast AR measure is a simple, easy-to-perform method for AR evaluation. It is particularly suitable for testing the effects of rapidly acting stimuli.