title: Amphiregulin promotes the proliferation of trophoblast cells during preimplantation development of porcine embryos.
abstract: Our objective was to investigate the effects of in vitro culture (IVC) medium supplemented with amphiregulin (AREG) on the preimplantation embryonic development of porcine (Genus: Sus domestica, Species: Landrace) embryos derived from in vitro fertilization (IVF) and parthenogenetic activation (PA). In vitro fertilization and PA embryos at the 1-cell stage were cultured in IVC medium supplemented with 0, 0.5, 5, or 50 ng/mL AREG for 7 d. There were significantly greater total numbers of cells in blastocysts of IVF and PA embryos cultured with 50 ng/mL AREG compared with that of controls. In vitro fertilization and PA embryos were then cultured in NCSU-23 medium supplemented with 50 ng/mL AREG on Days 1 through 7, Days 1 through 3 (early stage), or Days 4 through 7 (late stage), or without AREG. There were significantly greater numbers of trophoblast cells in the late-stage and full-term groups of IVF and PA embryos than in the early-stage and control groups. The presence of AREG protein in IVF-derived blastocysts was detected using a polyclonal AREG antibody and indirect immunofluorescence. Amphiregulin protein was localized in both the cytoplasm and nucleus. Using real-time polymerase chain reaction, we detected the expression of AREG mRNA in all developmental stages of IVF and PA embryos; however, the expression level varied according to stage. Thus, the incubation of porcine IVF and PA embryos in AREG-supplemented culture medium mainly at the late preimplantation stage increases the numbers of trophoblast cells.
authors.affiliation: College of Veterinary Medicine, Chungbuk National University, Cheongju 361-763, Korea.
authors.forename: J H
authors.initials: JH
authors.lastname: Lee
authors.forename: E
authors.initials: E
authors.lastname: Lee
authors.forename: D
authors.initials: D
authors.lastname: Biswas
authors.forename: C S
authors.initials: CS
authors.lastname: Jeung
authors.forename: G S
authors.initials: GS
authors.lastname: Lee
authors.forename: S H
authors.initials: SH
authors.lastname: Hyun
authors.forename: E B
authors.initials: EB
authors.lastname: Jeung
mesh_terms: D066258:Amphiregulin; D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D003470:Culture Media; D003593:Cytoplasm / Q000378:metabolism; D046149:Embryo Culture Techniques; D047108:Embryonic Development / Q000187:drug effects*; D005307:Fertilization in Vitro; D006023:Glycoproteins / Q000032:analysis / Q000378:metabolism / Q000494:pharmacology*; D036341:Intercellular Signaling Peptides and Proteins / Q000032:analysis / Q000378:metabolism / Q000494:pharmacology*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism; D013552:Swine; D014327:Trophoblasts / Q000187:drug effects* / Q000378:metabolism
chemical_list: D066258:Amphiregulin; D003470:Culture Media; D006023:Glycoproteins; D036341:Intercellular Signaling Peptides and Proteins; D012333:RNA, Messenger