title: Temporal expression of type I interferon receptor in the peri-implantation ovine extra-embryonic membranes: demonstration that human IFNalpha can bind to this receptor.
abstract: Interferon-tau (IFNtau), produced by the trophectoderm of ruminant ungulates, binds to the type I IFN receptor (IFNAR) located at the uterine endometrium in a paracrine manner. Since IFNtau attenuates the secretory pattern of an endometrial luteolysin, prostaglandin F2alpha, IFNtau has been considered as a conceptus factor implicated in the process of maternal recognition of pregnancy. Here we report the presence of IFNAR subunit (IFNAR1) in ovine conceptuses during the period of peri-implantation development and demonstrate that 125I-human (h) IFNalpha binds to membrane preparations from ovine corpus luteum and conceptus. Using an antibody against hIFNAR1, immunohistochemical analysis revealed that IFNAR1 protein was present in day 14 and 16 conceptuses (day 0 = day of estrus) and luminal and glandular epithelia of the endometrium. Conceptus membrane proteins analyzed by western blot with the same antibody displayed immunoreactive bands at 95, 60 and 55 kDa while endometrial membrane proteins showed bands at 200, 95 and 55 kDa. Northern blot analysis revealed that IFNAR1 mRNA was present in days 15-19 conceptuses and day 18-19 allantoic membranes. Receptor binding studies indicated that 125I-hIFNalpha binding to day 16, but not earlier, conceptus membrane proteins could be displaced with hIFNalpha or ovine IFNtau. Based on Scatchard analysis, day 16 conceptus membranes contained 28 fmol IFNAR/mg protein with a dissociation constant of 300 pM. Cross-linking experiments demonstrated that 125I-hIFNalpha-receptor complex migrated at 120 kDa, indicating that the receptor component(s) was approximately 100 kDa. These data provide evidence that although the binding does not occur until day 16, ovine conceptuses possess IFNAR1 near or at the time of implantation, suggesting that IFNtau, a factor produced by the trophectoderm of ruminant ungulates, could act on the conceptus in an autocrine manner. In addition to functioning as an antiluteolytic factor, therefore, IFNtau may have a direct effect on conceptus development.
authors.affiliation: Laboratory of Animal Breeding, Faculty of Agriculture, The University of Tokyo, Yayoi, Japan.
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Imakawa
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Tamura
authors.forename: Rita S F
authors.initials: RS
authors.lastname: Lee
authors.forename: Yongjie
authors.initials: Y
authors.lastname: Ji
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kogo
authors.forename: Senkiti
authors.initials: S
authors.lastname: Sakai
authors.forename: Ronald K
authors.initials: RK
authors.lastname: Christenson
mesh_terms: D000818:Animals; D015152:Blotting, Northern / Q000662:veterinary; D015153:Blotting, Western; D010064:Embryo Implantation / Q000276:immunology / Q000502:physiology*; D004717:Endometrium / Q000276:immunology / Q000378:metabolism; D005321:Extraembryonic Membranes / Q000276:immunology / Q000378:metabolism* / Q000502:physiology; D005260:Female; D007150:Immunohistochemistry / Q000662:veterinary; D016898:Interferon-alpha / Q000276:immunology / Q000378:metabolism* / Q000502:physiology; D008297:Male; D008565:Membrane Proteins; D011247:Pregnancy; D011270:Pregnancy, Animal / Q000276:immunology / Q000378:metabolism* / Q000502:physiology; D012313:RNA / Q000737:chemistry / Q000235:genetics; D053633:Receptor, Interferon alpha-beta; D017471:Receptors, Interferon / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D012756:Sheep / Q000276:immunology / Q000378:metabolism / Q000502:physiology*; D013997:Time Factors
chemical_list: C507964:IFNAR1 protein, human; D016898:Interferon-alpha; D008565:Membrane Proteins; D017471:Receptors, Interferon; D053633:Receptor, Interferon alpha-beta; D012313:RNA