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Production of motile acrosome‐reacted mouse sperm with nanomolar concentration of calcium ionophore A23187
Author(s) -
Tanphaichitr N.,
Hansen C.
Publication year - 1994
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/mrd.1080370312
Subject(s) - sperm , acrosome reaction , acrosome , ionophore , andrology , epididymis , biology , sperm motility , calcium , population , motility , bovine serum albumin , capacitation , immunology , microbiology and biotechnology , medicine , environmental health
A method to generate a population of motile, acrosome‐reacted mouse sperm is described. Sperm retrieved from the cauda epididymis and vas deferens were first capacited in a 3% bovine serum albumin (BSA) containing medium. Sperm were then resuspended in medium with low BSA content (0.01%) and treated with 30 nM of the calcium ionophore, A23187, which was added as a singel dose of 30 nM for 15 min at 37°C; or three sequential 10 nM doses over three 5 min intervals. Approximately 55–60% of the treated sperm population became acrosome reacted. The motility of the treated sperm sample was 40–65%, slightly lower than that of the control sperm, following addition of medium containing 3% BSA, This is in contrast to the <10% motility observed for capacitated mouse sperm treated with 10 μM A23187, a concentration that had been used by other investigators to induce the acrosome reaction. The ultrastructure of the 30 nM A23187‐induced acrosome‐reacted sperm ws similar to that of the acrosome‐reacted sperm induced by solubilized zonae pellucidae. These motile, acrosome‐reacted sperm were able to penetrate zone‐free mouse eggs at a higher rate than the control sperm. Thus this method of treatment will be useful for further physiological experimentation with acrosome‐reacted sperm. © 1994 Wiley‐Liss, Inc.