Heparin enhances protamine disulfide bond reduction during in vitro decondensation of human spermatozoa

Human Reproduction
Vanina Laura JulianelliJuan Carlos Calvo

Abstract

Human sperm nuclear decondensation in vivo involves protamine disulfide bond reduction by glutathione (GSH) and protamine/histone exchange, presumably with heparan sulfate (HS) as the protamine acceptor. The aim of the present study was to test the hypothesis that these two events occur simultaneously rather than sequentially, as has been hitherto accepted, and to test for the presence of HS in the human oocyte. Spermatozoa and isolated sperm nuclei obtained from normal volunteers were exposed in vitro to heparin, the functional analogue of HS and either GSH or dithiothreitol (DTT) as the disulfide reducing agent. Decondensing reagents were added either simultaneously or sequentially. Percentage sperm nuclear decondensation was assayed by phase contrast microscopy. Thiol reduced status of isolated sperm nuclei was evaluated both indirectly [acridine orange (AO) staining of acid-denatured DNA] and directly [monobromobimane (mBBr) staining of protamine-free thiols]. The presence of HS in mature metaphase II (MII) human oocytes was analyzed by immunocytochemistry. Sequential addition of reagents always resulted in significantly lower decondensation if GSH was used as the disulfide bond reducer (P < 0.05 for sperm and P < 0.001 for...Continue Reading

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Citations

Jul 18, 2014·Reproduction : the Official Journal of the Society for the Study of Fertility·Jingyu LiZhonghua Liu
Aug 31, 2013·The International Journal of Biochemistry & Cell Biology·Sun-Kyung LeeJoohong Ahnn
Aug 16, 2016·The Journal of Reproduction and Development·Tayita SuttirojpattanaMasaya Geshi
Nov 7, 2019·Journal of Assisted Reproduction and Genetics·C GalottoMarina Romanato

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