Genetic imprinting during impaired spermatogenesis.
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.
Hartmann
Sonja
S
Department of Urology and Pediatric Urology, Institute of Veterinary Anatomy, Histology and Embryology and Institute of Pathology, University of Giessen, Giessen Germany.
Bergmann
Martin
M
Bohle
Rainer M
RM
Weidner
Wolfgang
W
Steger
Klaus
K
D001324:Autoantigens / Q000235:genetics; D004247:DNA / Q000235:genetics / Q000378:metabolism; D019175:DNA Methylation; D018392:Genomic Imprinting / Q000235:genetics*; D006801:Humans; D007248:Infertility, Male / Q000235:genetics / Q000473:pathology; D008297:Male; D008957:Models, Genetic; D016133:Polymerase Chain Reaction; D062085:RNA, Long Noncoding; D022661:RNA, Untranslated / Q000235:genetics*; D017411:Ribonucleoproteins, Small Nuclear / Q000235:genetics; D013091:Spermatogenesis / Q000235:genetics*; D013094:Spermatozoa / Q000378:metabolism / Q000473:pathology; D055517:snRNP Core Proteins
D001324:Autoantigens; C094335:H19 long non-coding RNA; D062085:RNA, Long Noncoding; D022661:RNA, Untranslated; D017411:Ribonucleoproteins, Small Nuclear; C526417:SNRPN protein, human; D055517:snRNP Core Proteins; D004247:DNA