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Hum. Reprod. Advance Access published online on June 10, 2004
Human Reproduction, doi:10.1093/humrep/deh332
© 2004 by European Society of Human Reproduction and Embryology
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Received June 24, 2002
Revised March 18, 2004
Accepted April 30, 2004

Article

Studies on the membrane integrity of human sperm treated with a new injectable male contraceptive

K. Chaudhury 1, A.K. Bhattacharyya 2, S.K. Guha 1*

1 School of Medical Science and Technology, Indian Institute of Technology, Kharagpur 721302, India
2 Department of Biochemistry, University College of Science, University of Calcutta, Calcutta 700019, India

* To whom correspondence should be addressed. E-mail: guha_sk{at}yahoo.com.


  Abstract

BACKGROUND: The aim of this study was to evaluate the integrity of sperm surface characteristics in the presence of a new male contraceptive, RISUG [1 mg styrene maleic anhydride (SMA)/100 µl dimethylsulphoxide (DMSO) in 1 ml sperm solution]. METHODS: Progressively motile human sperm were treated in vitro with RISUG. The cells were analysed for the release of 5'-nucleotidase (5'-NT) (a plasma membrane marker) using 3 mmol/l 5'-AMP and 3 mmol/l {beta}-glycerophosphate as substrates. Hyaluronidase (an acrosomal membrane marker) was analysed using hyaluronic acid as a substrate. The contents of free and total acrosin, and % proacrosin (all acrosome markers) were assayed using 0.5 mmol/l {alpha}-N-benzoyl-L-arginine ethylester (BAEE). RESULTS: RISUG caused almost complete disintegration of the plasma membrane leading to significant (P<0.0001) release of 5'-NT into the surrounding media. Complete dissolution of the acrosome with concomitant vesiculation of the membrane system, as judged from the loss of hyaluronidase, was observed. Total acrosin content in the sperm was also reduced to almost 10%, and proacrosin dropped to 13.2% in the presence of RISUG in comparison to 90.2% in control (P<0.0001), indicating dispersion of acrosomal contents. CONCLUSION: Under in vitro conditions, RISUG, at a concentration of 1 mg SMA dissolved in 100 µl of DMSO, caused significant damage to the acrosome and its contents, indicating loss of functional ability of sperm.

Key words: acrosin, hyaluronidase, male contraceptive, 5'-nucleotidase, sperm membrane destabilization


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