Mol. Hum. Reprod. Advance Access originally published online on April 11, 2006
Molecular Human Reproduction 2006 12(6):407-411; doi:10.1093/molehr/gal040
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Genetic imprinting during impaired spermatogenesis
1Department of Urology and Pediatric Urology, 2Institute of Veterinary Anatomy, Histology and Embryology and 3Institute of Pathology, University of Giessen, Giessen Germany
4 To whom correspondence should be addressed at: Klinik und Poliklinik für Urologie und Kinderurologie, Rudolf-Buchheim-Strasse 7, 35385 Giessen, Germany. E-mail: klaus.steger{at}chiru.med.uni-giessen.de
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.
Key words: genetic imprinting/H19/impaired spermatogenesis/methylation-specific PCR/single-strand conformation polymorphism
Sonja Hartmann is a member of the DFG Research Training Group 533 Cell-Cell Interaction in Reproduction.
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