title: [Study of chromatin decondensation factors in human spermatozoids by flow cytometry].
abstract: To date, the mechanisms responsible for radical change of chromatin structure in male germ cells during fertilization are unclear. Evidence suggesting the existence of proteolytic nuclear enzymes in mature human spermatozoids are presented in this work. The possible role of these previously unknown proteases in decondensation of chromatin of spermatozoids in a fertilized ovum is discussed. Application of the flow cytometry technique has shown that treatment of human spermatozoid nuclei with SH-reagents leads not only to destruction of disulfide bonds between protamine molecules that is necessary for their effective utilization but also induces specific endogenous proteolytic activity that consequently results in rather fast decondensation of chromatin followed by proteolytic cleavage of nuclear proteins. A chromatin decondensation process can be almost totally blocked by serine protease inhibitors and components of seminal fluid. An original cytochemical approach of binding of fluorescently labeled protease inhibitor to the target of investigation has been used in order to visualize the localization of proteases in male germ cell nuclei. The results of our study suggest that one of the factors of chromatin reorganization involved in the formation of male pronucleus is endogenous nuclear protease of spermatozoids, which is activated by glutathione or other SH-components of ovum cytoplasm.
authors: E V
authors: EV
authors: Semenova
authors: M V
authors: MV
authors: Filatov
mesh_terms: D002467:Cell Nucleus / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism / Q000648:ultrastructure; D002843:Chromatin / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure*; D042003:DNA Packaging / Q000187:drug effects; D005434:Flow Cytometry; D006801:Humans; D008297:Male; D011480:Protease Inhibitors / Q000494:pharmacology; D013094:Spermatozoa / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism* / Q000648:ultrastructure; D013438:Sulfhydryl Compounds / Q000494:pharmacology
chemical_list: D002843:Chromatin; D011480:Protease Inhibitors; D013438:Sulfhydryl Compounds