Regulated expression of two sets of paternally imprinted genes is necessary for mouse parthenogenetic development to term.
Mouse parthenogenetic embryos (PEs) are developmentally arrested until embryo day (E) 9.5 because of genomic imprinting. However, we have shown that embryos containing genomes from non-growing (ng) and fully grown (fg) oocytes, i.e. ng(wt)/fg(wt) PE (wt, wild type), developed to E13.5. Moreover, parthenogenetic development could be extended to term by further regulation of Igf2 and H19 expression using mice with deletion of the H19 transcription unit (H19Delta13) together with its differentially unit (DMR). To gain an insight into the extended development of the parthenotes to term, we have here investigated the expression levels of paternally imprinted genes in ng(H19Delta13)/fg(wt) PE throughout their development. In ng(H19Delta13)/fg(wt) Pes that died soon after recovery, the expression of Igf2 and H19 was restored to the appropriate levels except for low Igf2 expression in the liver after E15.5. Further, the paternally expressed Dlk1 and Dio3 were repressed, while the expression levels of the maternal Gtl2 and Mirg were twice those of the controls. However, the above-mentioned four genes showed almost normal expression in the surviving ng(H19Delta13)/fg(wt) PEs. The methylation analysis revealed that the intragenic DMR of the Dlk1-Gtl2 domain was hypermethylated in the ng(H19Delta13)/fg(wt) PEs that survived, but not in the PEs that died soon after recovery. The present study suggests that two sets of co-ordinately regulated but oppositely expressed genes, Igf2-H19 and Dlk1-Gtl2, act as a critical barrier to parthenogenetic development in order to render a paternal contribution obligatory for descendants in mammals.
Wu
Qiong
Q
Department of BioScience, Tokyo University of Agriculture, Setagaya-ku, Tokyo 156-8502, Japan.
Kumagai
Takuya
T
Kawahara
Manabu
M
Ogawa
Hidehiko
H
Hiura
Hitoshi
H
Obata
Yayoi
Y
Takano
Riya
R
Kono
Tomohiro
T
D000818:Animals; D002135:Calcium-Binding Proteins; D019175:DNA Methylation; D047108:Embryonic Development / Q000502:physiology*; D005260:Female; D005313:Fetal Death / Q000235:genetics; D018507:Gene Expression Regulation, Developmental; D018392:Genomic Imprinting; D005865:Gestational Age; D007335:Insulin-Like Growth Factor II / Q000235:genetics; D036341:Intercellular Signaling Peptides and Proteins; D008297:Male; D008565:Membrane Proteins / Q000235:genetics; D051379:Mice; D010312:Parthenogenesis / Q000235:genetics* / Q000502:physiology; D011506:Proteins / Q000235:genetics; D062085:RNA, Long Noncoding; D012097:Repressor Proteins / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
D002135:Calcium-Binding Proteins; C512925:Dlk1 protein, mouse; D036341:Intercellular Signaling Peptides and Proteins; C422882:MEG3 non-coding RNA, mouse; D008565:Membrane Proteins; D011506:Proteins; D062085:RNA, Long Noncoding; D012097:Repressor Proteins; D007335:Insulin-Like Growth Factor II