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Hum. Reprod. Advance Access published online on June 18, 2007
Human Reproduction, doi:10.1093/humrep/dem166
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© The Author 2007. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

Motile human normozoospermic and oligozoospermic semen samples show a difference in double-strand DNA break incidence

Alwin A.H.A. Derijck1,2,*, Godfried W. van der Heijden1,3,*, Liliana Ramos1, Maud Giele1,4, Jan A.M. Kremer1 and Peter de Boer1,5

1 Department of Obstetrics and Gynaecology, Radboud University Nijmegen Medical Centre, PO Box 9101, 6500 HB Nijmegen, The Netherlands 2 Present address: University Medical Center Utrecht, Department of Pharmacology and Anatomy, Rudolf Magnus Institute of Neuroscience, Universiteitsweg 100, 3584 CG, Utrecht, The Netherlands 3 Present address: Carnegie Institution of Washington, Department of Embryology, 3520 San Martin Drive, Baltimore, MD 21218, USA 4 Present address: Orthopedic Research Laboratory, Department of Orthopedics, Radboud University Nijmegen Medical Centre, PO Box 9101, 6500 HB Nijmegen, The Netherlands

5 Correspondence address. Tel: +31-243610869; Fax: +31-24-3668597; E-mail: p.deboer{at}obgyn.umcn.nl

BACKGROUND: Among ICSI children de novo structural chromosome aberrations of male descent are increased. Misrepair of double-strand DNA breaks (DSBs) is a prerequisite for such aberrations to occur. To date, no absolute assessment of the number of DSBs in human sperm nuclei after gamete fusion has been described.

METHODS: Using man-mouse heterologous ICSI and {gamma}H2AX immunofluorescent staining, capable of detecting a single DSB, the number of lesions in ICSI selected sperm from normozoospermic men (n = 2) and oligozoospermic patients (n = 3) was quantified. A comparison with a subfertile male mouse model (n = 5) has been made. In addition, the fate of morphologically normal ejaculated immotile sperm after ICSI was examined.

RESULTS: A significant increase in the fraction of sperm cells bearing DSBs was found in oligozoospermic semen compared with that from normozoospermic men (P < 0.01). The majority of morphologically normal immotile human sperm showed excess {gamma}H2AX staining and nuclear disintegration. However, some had a non-deviant DSB pattern.

CONCLUSIONS: The increased fraction of DSB-positive sperm in both human and mouse oligozoospermic semen is adding to the surmise that semen from oligozoospermic patients has a reduced chromatin quality, causally related to reduced preimplantation embryo development. The use of ejaculated immotile sperm for in vitro reproduction is debatable due to sperm DNA degradation.

Key words: ICSI/double-strand DNA break/sperm/chromatin/DNA degradation

* Both authors contributed equally.

Submitted on November 13, 2006; resubmitted on April 16, 2007; accepted on May 15, 2007.


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