title: Determinants of orofacial clefting I: Effects of 5-Aza-2'-deoxycytidine on cellular processes and gene expression during development of the first branchial arch.
abstract: In this study, we identify gene targets and cellular events mediating the teratogenic action(s) of 5-Aza-2'-deoxycytidine (AzaD), an inhibitor of DNA methylation, on secondary palate development. Exposure of pregnant mice (on gestation day (GD) 9.5) to AzaD for 12h resulted in the complete penetrance of cleft palate (CP) in fetuses. Analysis of cells of the embryonic first branchial arch (1-BA), in fetuses exposed to AzaD, revealed: 1) significant alteration in expression of genes encoding several morphogenetic factors, cell cycle inhibitors and regulators of apoptosis; 2) a decrease in cell proliferation; and, 3) an increase in apoptosis. Pyrosequencing of selected genes, displaying pronounced differential expression in AzaD-exposed 1-BAs, failed to reveal significant alterations in CpG methylation levels in their putative promoters or gene bodies. CpG methylation analysis suggested that the effects of AzaD on gene expression were likely indirect.
authors.affiliation: Department of Molecular, Cellular and Craniofacial Biology, ULSD, University of Louisville, Louisville, KY 40202, USA. Electronic address: p0mukho1@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: 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: francine.rezzoug@louisville.edu.
authors.forename: Francine
authors.initials: F
authors.lastname: Rezzoug
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: guy.brock@louisville.edu.
authors.forename: Guy
authors.initials: G
authors.lastname: Brock
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; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D001374:Azacitidine / Q000031:analogs & derivatives* / Q000633:toxicity; D001934:Branchial Region / Q000187:drug effects* / Q000196:embryology / Q000473:pathology; D049109:Cell Proliferation / Q000187:drug effects; D002972:Cleft Palate / Q000139:chemically induced* / Q000196:embryology / Q000235:genetics / Q000473:pathology; 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; Apoptosis; Cleft palate; DNA methylation; Embryo; Proliferation