Induction of trophoblastic interferon expression in ovine blastocysts after treatment with double-stranded RNA.
Ovine trophoblast protein-1 (oTP-1) is an interferon (IFN) related to the IFN-omega. The objectives of this research were: (i) to attempt to induce oTP-1 mRNA in day-11 ovine conceptuses with polyinosinic-polycytidylic acid (poly(I).poly(C], and (ii) to determine if IFN-omega mRNA is also produced on day 11 of gestation. In experiment I, conceptuses were cultured in presence of 100 micrograms/ml poly(I).poly(C) (n = 5) or medium alone (control, n = 3) for up to 8 h. In situ hybridization was used to assess effects of treatment on mRNA concentrations for oTP-1 and actin (positive hybridization control). Poly(I).poly(C) increased oTP-1 mRNA concentrations approximately 2.5-fold (p less than 0.01), but had no effect on actin mRNA. In experiment II, the presence of mRNA for oTP-1 and ovine IFN-omega was determined by using reverse transcription-polymerase chain reaction (RT-PCR) analysis of conceptus total RNA coupled with Southern blot hybridization of the PCR reaction products with specific cDNA probes. oTP-1 mRNA was detectable in all poly(I).poly(C)-treated (n = 7) and control (n = 6) conceptuses, whereas IFN-omega mRNA was detected in only three of seven poly(I).poly(C)-treated conceptuses and not in any controls. Together these results demonstrate that expression of oTP-1 mRNA can be enhanced by treatment with poly(I).poly(C) and that oTP-1 is the primary but not the only type I-IFN inducible in conceptuses on day 11 of gestation.
Department of Animal Sciences, University of Missouri-Columbia 65211.
C E
CE
Farin
J C
JC
Cross
N A
NA
Tindle
C N
CN
Murphy
P W
PW
Farin
R M
RM
Roberts
D000818:Animals; D001483:Base Sequence; D001755:Blastocyst / Q000187:drug effects*; D015139:Blotting, Southern; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects*; D005865:Gestational Age; D007370:Interferon Type I / Q000235:genetics*; D008186:Luteolytic Agents / Q000037:antagonists & inhibitors; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D011070:Poly I-C / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011257:Pregnancy Proteins / Q000235:genetics*; D012330:RNA, Double-Stranded / Q000494:pharmacology*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000187:drug effects; D012756:Sheep
D007370:Interferon Type I; D008186:Luteolytic Agents; D011257:Pregnancy Proteins; D012330:RNA, Double-Stranded; D012333:RNA, Messenger; C020947:interferon tau; D011070:Poly I-C