title: Determinants of orofacial clefting II: Effects of 5-Aza-2'-deoxycytidine on gene methylation during development of the first branchial arch.
abstract: Defects in development of the secondary palate, which arise from the embryonic first branchial arch (1-BA), can cause cleft palate (CP). Administration of 5-Aza-2'-deoxycytidine (AzaD), a demethylating agent, to pregnant mice on gestational day 9.5 resulted in complete penetrance of CP in fetuses. Several genes critical for normal palatogenesis were found to be upregulated in 1-BA, 12h after AzaD exposure. MethylCap-Seq (MCS) analysis identified several differentially methylated regions (DMRs) in DNA extracted from AzaD-exposed 1-BAs. Hypomethylated DMRs did not correlate with the upregulation of genes in AzaD-exposed 1-BAs. However, most DMRs were associated with endogenous retroviral elements. Expression analyses suggested that interferon signaling was activated in AzaD-exposed 1-BAs. Our data, thus, suggest that a 12-h in utero AzaD exposure demethylates and activates endogenous retroviral elements in the 1-BA, thereby triggering an interferon-mediated response. This may result in the dysregulation of key signaling pathways during palatogenesis, causing CP.
authors.affiliation: Department of Molecular, Cellular and Craniofacial Biology, ULSD, University of Louisville, Louisville, KY 40202, USA. Electronic address: rsseel01@louisville.edu.
authors.forename: Ratnam S
authors.initials: RS
authors.lastname: Seelan
authors.affiliation: Department of Molecular, Cellular and Craniofacial Biology, ULSD, University of Louisville, Louisville, KY 40202, USA. Electronic address: p0mukh01@louisville.edu.
authors.forename: Partha
authors.initials: P
authors.lastname: Mukhopadhyay
authors.affiliation: Department of Molecular, Cellular and Craniofacial Biology, ULSD, University of Louisville, Louisville, KY 40202, USA. Electronic address: dwarn01@louisville.edu.
authors.forename: Dennis R
authors.initials: DR
authors.lastname: Warner
authors.affiliation: Department of Molecular, Cellular and Craniofacial Biology, ULSD, University of Louisville, Louisville, KY 40202, USA. Electronic address: irina.smolenkova@louisville.edu.
authors.forename: Irina A
authors.initials: IA
authors.lastname: Smolenkova
authors.affiliation: Department of Molecular, Cellular and Craniofacial Biology, ULSD, University of Louisville, Louisville, KY 40202, USA. Electronic address: drmikeky@gmail.com.
authors.forename: M Michele
authors.initials: MM
authors.lastname: Pisano
authors.affiliation: Department of Molecular, Cellular and Craniofacial Biology, ULSD, University of Louisville, Louisville, KY 40202, USA. Electronic address: dr.bob.greene@gmail.com.
authors.forename: Robert M
authors.initials: RM
authors.lastname: Greene
mesh_terms: D000818:Animals; D001374:Azacitidine / Q000031:analogs & derivatives* / Q000633:toxicity; D001934:Branchial Region / Q000187:drug effects* / Q000196:embryology; D002972:Cleft Palate / Q000139:chemically induced* / Q000196:embryology / Q000235:genetics; D019175:DNA Methylation / Q000187:drug effects*; D000077209:Decitabine; D047108:Embryonic Development / Q000187:drug effects* / Q000235:genetics; D005260:Female; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects*; D005865:Gestational Age; D008813:Mice, Inbred ICR; D011247:Pregnancy
chemical_list: D000077209:Decitabine; D001374:Azacitidine
keywords: 5-Aza-2′deoxycytidine; Cleft palate; DNA methylation; First branchial arch; MethylCap-Seq