title: LINE-1 retrotransposon copies are amplified during murine early embryo development.
abstract: Two large families of retrotransposons, that is, LINE-1 (Long Interspersed Nuclear Elements-1) and endogenous retroviruses, encode reverse transcriptase (RT) proteins in vertebrates. We previously showed that mouse preimplantation embryos are endowed with an endogenous, functional RT activity. Inhibiting that activity by microinjecting antisense oligonucleotides against a highly active LINE-1 family member in mouse oocytes blocked developmental progression between the two- and four-blastomere stages, indicating that LINE-1-encoded RT activity is strictly required at this critical transition in early development. Here we show that incubation of mouse zygotes with 5'-bromodeoxyuridine (BrdU) yields massive incorporation of this nucleoside analogue in newly synthesized DNA; surprisingly, a significant incorporation still occurs in both zygotic pronuclei in the presence of aphidicolin, a specific inhibitor of DNA replication. This aphidicolin-resistant BrdU incorporation is quantitatively abolished when embryos are simultaneously exposed to abacavir, a nucleoside RT inhibitor, indicating its retrotranscription-dependent nature. Moreover, quantitative PCR analysis revealed a burst of new synthesis of LINE-1 copies at the zygote- and two-cell embryo stages. These findings support the conclusion that RT-dependent amplification of LINE-1 retrotransposons is a distinctive feature of early embryonic genomes. Its physiological involvement in preimplantation murine development is discussed.
authors.affiliation: Italian National Institute of Health, Rome, Italy.
authors.forename: Patrizia
authors.initials: P
authors.lastname: Vitullo
authors.forename: Ilaria
authors.initials: I
authors.lastname: Sciamanna
authors.forename: Marta
authors.initials: M
authors.lastname: Baiocchi
authors.forename: Paola
authors.initials: P
authors.lastname: Sinibaldi-Vallebona
authors.forename: Corrado
authors.initials: C
authors.lastname: Spadafora
mesh_terms: D000818:Animals; D001973:Bromodeoxyuridine / Q000493:pharmacokinetics; D002467:Cell Nucleus / Q000378:metabolism; D002970:Cleavage Stage, Ovum / Q000378:metabolism / Q000502:physiology; D056915:DNA Copy Number Variations / Q000502:physiology; D004261:DNA Replication / Q000235:genetics; D004622:Embryo, Mammalian; D047108:Embryonic Development / Q000235:genetics*; D005260:Female; D005784:Gene Amplification / Q000502:physiology; D018507:Gene Expression Regulation, Developmental; D020084:Long Interspersed Nucleotide Elements / Q000235:genetics*; D008297:Male; D051379:Mice; D018626:Retroelements / Q000235:genetics; D015053:Zygote / Q000378:metabolism
chemical_list: D018626:Retroelements; D001973:Bromodeoxyuridine