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Mol. Hum. Reprod. Advance Access published online on April 11, 2006
Molecular Human Reproduction, doi:10.1093/molehr/gal040
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© The Author 2006. 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 The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions@oxfordjournals.org
Received February 16, 2006
Accepted March 17, 2006

Article

Genetic imprinting during impaired spermatogenesis

Sonja Hartmann 1, Martin Bergmann 2, Rainer M. Bohle 3, Wolfgang Weidner 4, and Klaus Steger 1 *

1 Department of Urology and Pediatric Urology, University of Giessen, Giessen, Germany
2 Institute of Veterinary Anatomy, Histology and Embryology, University of Giessen, Giessen, Germany
3 Institute of Pathology, University of Giessen, Giessen, Germany
4 Department of Urology and Pediatric Urology, University of Giessen, Giessen Germany

* To whom correspondence should be addressed.
Klaus Steger, E-mail: klaus.steger{at}chiru.med.uni-giessen.de


  Abstract

Disorders in genetic imprinting are discussed as potential genetic risk in assisted reproduction technology (ART), where most of the natural selection mechanisms are bypassed. As currently only limited information about genomic imprinting in disruptive spermatogenesis is available, we analysed the imprinting state of the paternally methylated gene H19 in various germ cell populations derived from seminiferous tubules exhibiting impaired spermatogenesis. Different germ cell types were isolated by laser microdissection from human testicular paraffin sections. Although the methylation state of the maternally imprinted gene SNRPN was investigated by methylation-specific PCR (M-PCR) to establish the isolation method, methylation of H19 was analysed by a single-strand conformation-based method. Contamination by somatic Sertoli cells was excluded because of Sertoli cell-specific vimentin immunohistochemistry before germ cell laser microdissection. We demonstrate correct genetic imprints for H19 even in spermatogonia selected from seminiferous tubules exhibiting spermatogenic arrest at the level of spermatogonia, providing no evidence for incorrect genomic imprinting in spermatozoa from infertile men used for ICSI.

Keywords: genetic imprinting/H19/impaired spermatogenesis/methylation-specific PCR/single-strand conformation polymorphism.
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