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Human Reproduction, Vol. 11, No. 4, pp. 741-745, 1996
© 1996 European Society of Human Reproduction and Embryology

research-article

A comparison of three methods for detecting the acrosome reaction in human spermatozoa*

Abdel-Halim Amin1, Janice L. Bailey2, Bayard T. Storey, Luis Blasco and Susan Heyner3

Division of Reproductive Biology, Department of Obstetrics and Gynecology, University of Pennsylvania Medical Center 311D John Morgan Building, 3620 Hamilton Walk, Philadelphia, PA 19104-6080, USA

Correspondence: 3To wgom corespondence should be addressed

This study was designed to compare three different fluorescent probes to assay the acrosome reaction in human spermatozoa: chlortetracycline (CTC), mannosylated bovine serum albumin (BSA) labelled with fluorescein (MAF), and quinacrine (QN)- Normal human sperm ejaculates were washed and allowed to swim up for 30–60 min. Samples were examined under epifluorescence for the percentage of the acrosome reacted spermatozoa, as detected by the three probes. There was no significant difference between samples of fresh, uncapadtated spermatozoa evaluated with CTC, MAF or QN; all gave <10% reacted. Following capacitation for 3 h, the percentage of spontaneously reacted spermatozoa was higher than in fresh spermatozoa; CTC and MAF gave the same percentage (12%), while QN indicated a higher percentage (18%) of reacted spermatozoa (P < 0.001). Following exposure to ionophore A23187 at 1 h, the percentage of acrosome reactions increased to a mean of 31% as detected with CTC or MAF; the mean percentage (45%) was significantly higher with QN (P < 0.0001). Further incubation up to 2 h with A23187 did not change these percentages. These results suggest that the QN probe detects the onset stage of the acrosome reaction, whereas the CTC and MAF probes detect the later stages in which the acrosomal cap is lost. Use of the two types of probe provides a means for finer resolution of the time course of the acrosome reaction in the human spermatozoa.

Key words: acrosome reaction/chlortetracycline/human spermatozoa/mannosylated BSA/quinacrine

*This work was supported in part by NIH grant HD-06274 and by a grant from the Egyptian Government to A.-H.Amin.

1Visiting Scholar from Minia University, Minia, Egypt

2Present address: Département des sciences animales, Université Laval, Pavillon Pierre-Comtois, Ave. de 1'Agriculture, Ste.-Foy, Québec, PQ, Canada G1K 7P4


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