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Human Molecular Genetics, 2000, Vol. 9, No. 14 2183-2187
© 2000 Oxford University Press

Establishment of the paternal methylation imprint of the human H19 and MEST/PEG1 genes during spermatogenesis

Antoine Kerjean1,3, Jean-Michel Dupont2, Christian Vasseur3, Dominique Le Tessier2, Laurence Cuisset3, Andràs Pàldi4,+, Pierre Jouannet1 and Marc Jeanpierre3

1Laboratoire de Biologie de la Reproduction, 2Laboratoire d’Histologie Embryologie Cytogénétique and 3Laboratoire de Biochimie et Génétique Moléculaire, Université Paris V, Hôpital Cochin-Port-Royal, Pavillon Cassini, 123 Boulevard de Port-Royal, 75014 Paris, France and 4INSERM U257, Institut Cochin de Génétique Moléculaire (ICGM), Centre Hospitalier Universitaire-Cochin, 24 rue du Faubourg Saint-Jacques, 75014 Paris, France

Parental-specific epigenetic modifications are imprinted on a subset of genes in the mammalian genome during germ cell maturation. However, the precise timing of their establishment remains to be determined. Methylation of CpG dinucleotides has been shown to be a part of the parental imprint. We have examined how the methylation pattern characteristic of the paternal allele in germ cells are established during human spermatogenesis. Two representative imprinted genes, H19 and MEST/PEG1, were studied. The experiments were performed using the bisulphite sequencing method on microdissected individual cells at different stages of male germ cell differentiation. We show that both genes are unmethylated in fetal spermatogonia, suggesting that all pre-existing methylation imprints are already erased by this stage. The MEST/PEG1 gene remains unmethylated at all subsequent post-pubertal stages of spermatogenesis, including mature spermatozoa. The methylation of H19 typical of the paternal allele first appears in a subset of adult spermatogonia and then is maintained in spermatocytes, spermatids and mature spermatozoa. Our results suggest that the methylation imprint inherited from the parents is first erased in the male germ line at an early fetal stage. The paternal-specific imprint is re-established only later, during spermatogonial differentiation in the adult testis.

+ To whom correspondence should be addressed. Tel: +33 1 44 41 24 56; Fax: +33 1 44 41 24 62; Email: paldi@cochin.inserm.fr


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