title: Induction of autophagy improves embryo viability in cloned mouse embryos.
abstract: Autophagy is an essential cellular mechanism that degrades cytoplasmic proteins and organelles to recycle their components. Moreover, autophagy is essential for preimplantation development in mammals. Here we show that autophagy is also important for reprogramming in somatic cell nuclear transfer (SCNT). Our data indicate that unlike fertilized oocytes, autophagy is not triggered in SCNT embryos during 6 hours of activation. Mechanistically, the inhibited autophagic induction during SCNT activation is due to the cytochalasin B (CB) caused depolymerization of actin filaments. In this study, we induced autophagy during SCNT activation by rapamycin and pp242, which could restore the expected level of autophagy and significantly enhance the development of SCNT embryos to the blastocyst stage when compared with the control (68.5% and 68.7% vs. 41.5%, P < 0.05). Furthermore, the treatment of rapamycin and pp242 accelerates active DNA demethylation indicated by the conversion of 5 mC to 5 hmC, and treatment of rapamycin improves degradation of maternal mRNA as well. Thus, our findings reveal that autophagy is important for development of SCNT embryos and inhibited autophagic induction during SCNT activation might be one of the serious causes of low efficiency of SCNT.
authors.affiliation: Department of Histology and Embryology, Harbin Medical University, Harbin, China.
authors.forename: XingHui
authors.initials: X
authors.lastname: Shen
authors.affiliation: Department of Histology and Embryology, Harbin Medical University, Harbin, China.
authors.forename: Na
authors.initials: N
authors.lastname: Zhang
authors.affiliation: Department of Histology and Embryology, Harbin Medical University, Harbin, China.
authors.forename: ZhenDong
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Department of Histology and Embryology, Harbin Medical University, Harbin, China.
authors.forename: GuangYu
authors.initials: G
authors.lastname: Bai
authors.affiliation: Department of Histology and Embryology, Harbin Medical University, Harbin, China.
authors.forename: Zhong
authors.initials: Z
authors.lastname: Zheng
authors.affiliation: Department of Histology and Embryology, Harbin Medical University, Harbin, China.
authors.forename: YanLi
authors.initials: Y
authors.lastname: Gu
authors.affiliation: Department of Histology and Embryology, Harbin Medical University, Harbin, China.
authors.forename: YanShuang
authors.initials: Y
authors.lastname: Wu
authors.affiliation: Department of Histology and Embryology, Harbin Medical University, Harbin, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Liu
authors.affiliation: Department of Histology and Embryology, Harbin Medical University, Harbin, China.
authors.forename: DongJie
authors.initials: D
authors.lastname: Zhou
authors.affiliation: Department of Histology and Embryology, Harbin Medical University, Harbin, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Lei
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism; D000818:Animals; D001343:Autophagy / Q000187:drug effects / Q000235:genetics; D019976:Cloning, Organism; D019175:DNA Methylation; D004622:Embryo, Mammalian; D005260:Female; D005307:Fertilization in Vitro; D015870:Gene Expression; D008297:Male; D000076222:Mechanistic Target of Rapamycin Complex 1; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism; D046912:Multiprotein Complexes / Q000037:antagonists & inhibitors; D053652:Nuclear Transfer Techniques; D009865:Oocytes / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D020871:RNA Stability / Q000187:drug effects; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020123:Sirolimus / Q000494:pharmacology; D058570:TOR Serine-Threonine Kinases / Q000037:antagonists & inhibitors
chemical_list: C480860:Map1lc3b protein, mouse; D008869:Microtubule-Associated Proteins; D046912:Multiprotein Complexes; D047428:Protein Kinase Inhibitors; D012333:RNA, Messenger; D000076222:Mechanistic Target of Rapamycin Complex 1; D058570:TOR Serine-Threonine Kinases; D020123:Sirolimus