title: Selective expression of sense and antisense transcripts of the sushi-ichi-related retrotransposon--derived family during mouse placentogenesis.
abstract: BACKGROUND
  LTR-retrotransposons became functional neogenes through evolution by acquiring promoter sequences, regulatory elements and sequence modification. Mammalian retrotransposon transcripts (Mart1-9), also called sushi-ichi-related retrotransposon-homolog (SIRH) genes, are a class of Ty3/gypsy LTR-retroelements showing moderate homology to the sushi-ichi LTR-retrotransposon in pufferfish. Rtl1/Mart1 and Peg10/Mart2 expression in mouse placenta and demonstration of their functional roles during placental development exemplifies their importance in cellular processes. In this study, we analyzed all eleven mouse Mart genes from the blastocyst stage and throughout placentogenesis in order to gain information about their expression and regulation.
  
  
  RESULTS
  Quantitative PCR, in situ hybridization (ISH) and immunoblotting showed various expression patterns of the 11 mouse Mart genes through different placental stages. Zcchc5/Mart3, Zcchc16/ Mart4 and Rgag1/Mart9 expression was undetectable. Rtl1/Mart1, Peg10/Mart2, Rgag4/Mart5 - Cxx1a,b,c/Mart8b,c,a gene expression was very low at the blastocyst stage. Later placental stages showed an increase of expression for Rtl1/Mart1, Rgag4/Mart5 - Cxx1a,b,c/Mart8b,c,a, the latter up to 1,489 molecules/ng cDNA at E9.5. From our recently published findings Peg10/Mart2 was the most highly expressed Mart gene. ISH demonstrated sense and antisense transcript co-localization of Rgag4/Mart5 to Cxx1a,b,c/Mart8b,c,a in trophoblast subtypes at the junctional zone, with an accumulation of antisense transcripts in the nuclei. To validate these results, we developed a TAG-aided sense/antisense transcript detection (TASA-TD) method, which verified sense and antisense transcripts for Rtl1/Mart1, Rgag4/Mart5 - Cxx1a,b,c/Mart8b,c,a. Except for Rtl1/Mart1 and Cxx1a,b/Mart8b,c all other Mart genes showed a reduced amount of antisense transcripts. Northern blot and 5' and 3' RACE confirmed both sense and antisense transcripts for Ldoc1/Mart7 and Cxx1a,b,c/Mart8b,c,a. Immunoblotting demonstrated a single protein throughout all placental stages for Ldoc1/Mart7, but for Cxx1a,b,c/Mart8b,c,a a switch occurred from a 57 kDa protein at E10.5 and E14.5 to a 25 kDa protein at E16.5 and E18.5.
  
  
  CONCLUSIONS
  RNA and protein detection of mouse Mart genes support neo-functionalization of retrotransposons in mammalian genomes. Undetectable expression of Zcchc5/Mart3, Zcchc16/Mart4 and Rgag1/Mart9 indicate no role during mouse placentogenesis. Rgag4/Mart5 to Cxx1a,b,c/Mart8b,c,a gene expression support a role for differentiation from the ectoplacental cone. Mart antisense transcripts and protein alterations predict unique and complex molecular regulation in a time directed manner throughout mouse placentogenesis.
authors.affiliation: Department of Gynaecology and Obstetrics, Laboratory for Molecular Medicine, Friedrich-Alexander University Erlangen-Nürnberg (FAU), University-Clinic Erlangen, Erlangen, Germany. Christine.Henke@uk-erlangen.de.
authors.forename: Christine
authors.initials: C
authors.lastname: Henke
authors.affiliation: Department of Gynaecology and Obstetrics, Laboratory for Molecular Medicine, Friedrich-Alexander University Erlangen-Nürnberg (FAU), University-Clinic Erlangen, Erlangen, Germany. strisspa@outlook.com.
authors.forename: Pamela L
authors.initials: PL
authors.lastname: Strissel
authors.affiliation: Department of Gynaecology and Obstetrics, Laboratory for Molecular Medicine, Friedrich-Alexander University Erlangen-Nürnberg (FAU), University-Clinic Erlangen, Erlangen, Germany. Theresa.Schubert@uk-erlangen.de.
authors.forename: Maria-Theresa
authors.initials: MT
authors.lastname: Schubert
authors.affiliation: Department of Gynaecology and Obstetrics, Laboratory for Molecular Medicine, Friedrich-Alexander University Erlangen-Nürnberg (FAU), University-Clinic Erlangen, Erlangen, Germany. mogmitchell@gmail.com.
authors.forename: Megan
authors.initials: M
authors.lastname: Mitchell
authors.affiliation: Institute of Biochemistry, D-91054, Erlangen, Germany. ccstolt@biochem.uni-erlangen.de.
authors.forename: Claus C
authors.initials: CC
authors.lastname: Stolt
authors.affiliation: Department of Gynaecology and Obstetrics, Laboratory for Molecular Medicine, Friedrich-Alexander University Erlangen-Nürnberg (FAU), University-Clinic Erlangen, Erlangen, Germany. Florian.Faschingbauer@uk-erlangen.de.
authors.forename: Florian
authors.initials: F
authors.lastname: Faschingbauer
authors.affiliation: Department of Gynaecology and Obstetrics, Laboratory for Molecular Medicine, Friedrich-Alexander University Erlangen-Nürnberg (FAU), University-Clinic Erlangen, Erlangen, Germany. fk-direktion@uk-erlangen.de.
authors.forename: Matthias W
authors.initials: MW
authors.lastname: Beckmann
authors.affiliation: Department of Gynaecology and Obstetrics, Laboratory for Molecular Medicine, Friedrich-Alexander University Erlangen-Nürnberg (FAU), University-Clinic Erlangen, Erlangen, Germany. Reiner.Strick@uk-erlangen.de.
authors.forename: Reiner
authors.initials: R
authors.lastname: Strick
mesh_terms: D000818:Animals; D005260:Female; D015870:Gene Expression; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D010920:Placenta / Q000196:embryology*; D011247:Pregnancy; D016372:RNA, Antisense / Q000096:biosynthesis*; D012333:RNA, Messenger / Q000096:biosynthesis*; D060888:Real-Time Polymerase Chain Reaction; D018626:Retroelements; D014158:Transcription, Genetic
chemical_list: D016372:RNA, Antisense; D012333:RNA, Messenger; D018626:Retroelements