title: Poly(ADP-ribosyl)ation is involved in pro-survival autophagy in porcine blastocysts.
abstract: Poly(ADP-ribosyl)ation (PARylation) prevents apoptosis through its involvement in pro-survival autophagy in cultured cells; whether or not the same is true for pre-implantation embryos has not yet been documented. In this study, we investigated the participation of PARylation and autophagy in in vitro porcine pre-implantation embryo development. The transcript levels of autophagy-related genes and poly(ADP-ribose) polymerase 1 (PARP1), an enzyme required for PARylation, were transiently up-regulated by fertilization, decreased at the late 1-cell stage, and maintained until the blastocyst stage. LC3, a marker of autophagosomes, and poly(ADP-ribose) (PAR) polymer were present in all stages of pre-implantation development. Exposure of embryos to 3-methyladenine, an autophagy inhibitor, or 3-aminobenzamide, a PARP inhibitor, suppressed the development of blastocysts. Pharmacological inhibition of PARylation further suppressed pro-survival autophagy by decreasing the expression of autophagy-related genes (ATG5, BECLIN1, and LC3) and decreasing LC3 protein abundance while increasing the rate of apoptosis in blastocysts. Deficiency in autophagy also induced abnormal accumulation of SQSTM1/p62 aggregates in porcine blastocysts. Collectively, these data suggest that PARylation is involved in selective autophagic degradation of ubiquitinated proteins, functioning in a pro-survival role, in porcine in vitro-produced embryos. These pro-survival regulatory mechanisms may be important for the control of embryo quality.
authors.affiliation: Department of Animal Biotechnology, Bio-Organ Research Center/Animal Resources Research Center, Konkuk University, Seoul, South Korea.
authors.forename: Hye Ran
authors.initials: HR
authors.lastname: Lee
authors.affiliation: Department of Biotechnology and Medical Engineering, National Institute of Technology, Rourkela, Odisha, India.
authors.forename: Mukesh Kumar
authors.initials: MK
authors.lastname: Gupta
authors.affiliation: Department of Animal Biotechnology, Bio-Organ Research Center/Animal Resources Research Center, Konkuk University, Seoul, South Korea.
authors.forename: Duk Hyeon
authors.initials: DH
authors.lastname: Kim
authors.affiliation: Department of Animal Biotechnology, Bio-Organ Research Center/Animal Resources Research Center, Konkuk University, Seoul, South Korea.
authors.forename: Jeong Ho
authors.initials: JH
authors.lastname: Hwang
authors.affiliation: Department of Animal Biotechnology, Bio-Organ Research Center/Animal Resources Research Center, Konkuk University, Seoul, South Korea.
authors.forename: Bumsup
authors.initials: B
authors.lastname: Kwon
authors.affiliation: Department of Animal Biotechnology, Bio-Organ Research Center/Animal Resources Research Center, Konkuk University, Seoul, South Korea.
authors.forename: Hoon Taek
authors.initials: HT
authors.lastname: Lee
mesh_terms: D000818:Animals; D051017:Apoptosis Regulatory Proteins / Q000235:genetics / Q000378:metabolism; D001343:Autophagy / Q000235:genetics / Q000502:physiology*; D001755:Blastocyst / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D002470:Cell Survival / Q000235:genetics; D047108:Embryonic Development / Q000235:genetics; D005306:Fertilization / Q000235:genetics; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D011064:Poly Adenosine Diphosphate Ribose / Q000378:metabolism*; D011065:Poly(ADP-ribose) Polymerases / Q000502:physiology*; D013552:Swine
chemical_list: D051017:Apoptosis Regulatory Proteins; D011064:Poly Adenosine Diphosphate Ribose; D011065:Poly(ADP-ribose) Polymerases
keywords: PARylation; Poly-ADP ribosylation; autophagy; blastocyst; porcine