title: Induction of IFNT-Stimulated Genes by Conceptus-Derived Exosomes during the Attachment Period.
abstract: Biochemical and/or physical communication between the conceptus and the uterine endometrium is required for conceptus implantation to the maternal endometrium, leading to placentation and the establishment of pregnancy. We previously reported that in vitro co-culture system with bovine trophoblast CT-1 cells, primary uterine endometrial epithelial cells (EECs), and uterine flushings (UFs) mimics in vivo conceptus attachment process. To identify molecules in UFs responsible for this change, we first characterized protein contents of UFs from day 17 cyclic (C17) and pregnant (P17) ewes through the use of two dimensional-Polyacrylamide Gel Electrophoresis (2D-PAGE), followed by Liquid Chromatography-tandem Mass Spectrometry (LC-MS/MS) analysis. These analyses identified 266 proteins specific for P17 UFs, from which 172 proteins were identified as exosomal proteins. Among 172 exosomal proteins, 8 proteins that had been identified as exosomal proteins were chosen for further analysis, including macrophage-capping protein (CAPG), aldo-keto reductase family 1, member B1 protein (AKR1B1), bcl-2-like protein 15 (BCL2L15), carbonic anhydrase 2 (CA2), isocitrate dehydrogenase 2 (IDH2), eukaryotic translation elongation factor 2 (EEF2), moesin (MSN), and ezrin (EZR). CAPG and AKR1B1 were again confirmed in P15 and P17 UFs, and more importantly CAPG and AKR1B1, mRNA and protein, were found only in P15 and P17 conceptuses. Moreover, exosomes were isolated from C15, C17, P15, or P17 UFs. Only P15 and P17 exosomes, originated from the conceptus, contained interferon tau (IFNT) as well as CAPG and AKR1B1, and up-regulated STAT1, STAT2, MX1, MX2, BST2, and ISG15 transcripts in EECs. These observations indicate that in addition to endometrial derived exosomes previously described, conceptus-derived exosomes are present in UFs and could function to modify endometrial response. These results suggest that exosomes secreted from conceptuses as well as endometria are involved in cell to cell interactions for conceptus implantation to the maternal endometrium.
authors.affiliation: Animal Resource Science Center, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Ibaraki, Japan.|Department of Food, Agriculture and Environment, Miyagi University, Miyagi, Japan.
authors.forename: Keigo
authors.initials: K
authors.lastname: Nakamura
authors.affiliation: Laboratory of Theriogenology and Animal Breeding, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan.
authors.forename: Kazuya
authors.initials: K
authors.lastname: Kusama
authors.affiliation: Laboratory of Theriogenology and Animal Breeding, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan.
authors.forename: Rulan
authors.initials: R
authors.lastname: Bai
authors.affiliation: Department of Occupational and Environmental Health, Faculty of Pharmaceutical Sciences, Tokyo University of Science, Chiba, Japan.
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Sakurai
authors.affiliation: Department of Kampo Pharmacy, Yokohama University of Pharmacy, Kanagawa, Japan.
authors.forename: Kazuto
authors.initials: K
authors.lastname: Isuzugawa
authors.affiliation: Department of Animal Sciences, The University of Tennessee, Knoxville, TN, United States of America.
authors.forename: James D
authors.initials: JD
authors.lastname: Godkin
authors.affiliation: Department of Food, Agriculture and Environment, Miyagi University, Miyagi, Japan.
authors.forename: Yoshihito
authors.initials: Y
authors.lastname: Suda
authors.affiliation: Animal Resource Science Center, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Ibaraki, Japan.
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Imakawa
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D010064:Embryo Implantation; D004717:Endometrium / Q000378:metabolism* / Q000502:physiology; D055354:Exosomes / Q000378:metabolism*; D005260:Female; D007370:Interferon Type I / Q000378:metabolism*; D011247:Pregnancy; D011257:Pregnancy Proteins / Q000378:metabolism*; D020543:Proteome / Q000378:metabolism; D012756:Sheep
chemical_list: D007370:Interferon Type I; D011257:Pregnancy Proteins; D020543:Proteome; C020947:interferon tau