Study of chromatin decondensation factors in human spermatozoids by flow cytometry |
| |
Authors: | E V Semenova M V Filatov |
| |
Institution: | 1.Division of Molecular and Radiation Biophysics, Konstantinov Petersburg Nuclear Physics Institute,Russian Academy of Sciences,Gatchina, Leningrad oblast, Orlova Roshcha,Russia |
| |
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. |
| |
Keywords: | |
本文献已被 SpringerLink 等数据库收录! |
|