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Sperm decondensation during fertilisation in the mouse: presence of DNase I hypersensitive sites in situ and a putative role for topoisomerase II

Published online by Cambridge University Press:  13 November 2000

Davide Bizzaro ,
Giancarlo Manicardi ,
Patrizia Grace Bianchi  and
Denny Sakkas
Davide Bizzaro
Affiliation:
Department of Animal Biology, University of Modena and Reggio Emilia, Via Berengario 14, 41100 Modena, Italy.
Giancarlo Manicardi
Affiliation:
Department of Animal Biology, University of Modena and Reggio Emilia, Via Berengario 14, 41100 Modena, Italy.
Patrizia Grace Bianchi
Affiliation:
Clinic of Sterility, Department of Obstetrics and Gynaecology, University Hospital of Geneva, 20 rue Alcide Jentzer, 1211 Geneva 14, Switzerland.
Denny Sakkas
Affiliation:
Assisted Conception Unit, Birmingham Women's Hospital, Birmingham B15 2TG, UK.
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Abstract

In this study our aim was to characterise the presence and the role of DNA alterations during sperm decondensation in the mouse. To visualise the changes during decondensation we investigated for the presence of DNase I hypersensitive sites in situ and for a putative role for topoisomerase II by examining the effect of teniposide, a topoisomerase II inhibitor, during fertilisation. In situ nick translation without the previous addition of DNase I failed to reveal the presence of endogenous nicks in decondensing sperm and pronuclei whereas preincubation of fixed oocytes with DNase I indicated that decondensing sperm were sensitive to this enzyme. Addition of 100 μM teniposide did not completely inhibit pronuclei formation but its addition to the fertilisation medium did lead to the presence of endogenous DNA nicks in decondensing sperm. These observations suggest that DNase I hypersensitivity during sperm decondensation is related to the dramatic conformational changes that the chromatin undergoes during the decondensation process, in which topoisomerase II may be implicated.

Keywords

DNase IFertilisationIn situ nick translationSperm decondensationTopoisomerase II
Type
Research Article
Information
Zygote , Volume 8 , Issue 3 , August 2000 , pp. 197 - 202
Copyright
2000 Cambridge University Press

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