A Simple, Clinical Assay to Evaluate The Acrosin Activity of Human Spermatozoa

Wiley - Tập 10 Số 3 - Trang 221-231 - 1989
William P. Kennedy1, J. M. Kaminski1, H.H. van der Ven2, Rajasingam S. Jeyendran1,3, D Reid1, Jane M. Blackwell1, P. Bielfeld1, Lourens J.D. Zaneveld1
1Departments of Obstetrics and Gynecology, and Biochemistry Rush University, Rush-Presbyterian-St. Luke's Medical Center, Chicago, IL
2Department of Obstetrics and Gynecology, University of Bonn, Bonn, Germany
3Institute of Reproductive Medicine, Chicago, IL

Tóm tắt

Acrosin, a sperm‐specific arosomal proteinase, has an essential role in the fertilization process. Low levels of acrosin appear to be associated with subfertility and infertility, and the acrosin activity of spermatozoa may potentially be a useful indicator of semen quality. The standard acrosin tests employed by research laboratories are too complicated and/or time consuming for clinical use; therefore, a simple assay has been developed to assess total acrosin activity (acrosin and activatable proacrosin). To perform the test, liquefied semen is centrifuged over Ficoll, the washed sperm pellet is suspended in a detergent (Triton X‐100)‐substrate (N‐α‐benzoyl‐DL‐arginine p‐nitroanilide) buffer, pH. 8.0, and the amidase activity is determined spectrophotometrically after a 3‐hour incubation period. Amidase activity can be inhibited with benzamidine, indicating that the activity is primarily or entirely due to acrosin. The absence of detergent in the incubation medium results in greatly reduced activity. The assay is repeatable, linear with increasing sperm concentration, sensitive to a lower limit of 2 × 106 spermatozoa, and the results correspond to those obtained with a standard acrosin extraction and assay technique. Storage of ejaculates at 3 to 6 C or at 22 to 24 C for 24 hours does not affect the acrosin activity significantly but much higher temperatures can cause a loss of activity. Freezing ejaculates results in a large decrease in sperm acrosin activity. Leukocytes show minimal activity in the assay. Sperm populations prepared by a swim‐up procedure average approximately a 2‐fold higher acrosin activity than the original ejaculates. Preliminary experiments indicate that the average sperm acrosin activity of ejaculates whose spermatozoa successfully fertilize human eggs in vitro is significantly higher than those that do not fertilize eggs.

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Wiley

Tập 10 Số 3

221-231

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