title: Myocardial smad4 is essential for cardiogenesis in mouse embryos.
abstract: Congenital heart diseases are the most commonly observed human birth defects and are the leading cause of infant morbidity and mortality. Accumulating evidence indicates that transforming growth factor-beta/bone morphogenetic protein signaling pathways play critical roles during cardiogenesis. Smad4 encodes the only common Smad protein in mammals, which is a critical nuclear mediator of transforming growth factor-beta/bone morphogenetic protein signaling. The aim of this work was to investigate the roles of Smad4 during heart development. To overcome the early embryonic lethality of Smad4(-/-) mice, we specifically disrupted Smad4 in the myocardium using a Cre/loxP system. We show that myocardial-specific inactivation of Smad4 caused heart failure and embryonic lethality at midgestation. Histological analysis revealed that mutant mice displayed a hypocellular myocardial wall defect, which is likely the primary cause for heart failure. Both decreased cell proliferation and increased apoptosis contributed to the myocardial wall defect in mutant mice. Data presented in this article contradict a previous report showing that Smad4 is dispensable for heart development. Our further molecular characterization showed that expression of Nmyc and its downstream targets, including cyclin D1, cyclin D2, and Id2, were downregulated in mutant embryos. Reporter analysis indicated that the transcriptional activity of the 351-bp Nmyc promoter can be positively regulated by bone morphogenetic protein stimulation and negatively regulated by transforming growth factor-beta stimulation. Chromatin immunoprecipitation analysis revealed that the Nmyc promoter can form a complex with Smad4, suggesting that Nmyc is a direct downstream target of Smad4. In conclusion, this study provides the first mouse model showing that Smad4 plays essential roles during cardiogenesis.
authors.affiliation: Department of Genetics, Division of Genetic and Translational Medicine, The University of Alabama at Birmingham, Birmingham, AL 35294, USA.
authors.forename: Lanying
authors.initials: L
authors.lastname: Song
authors.forename: Wensheng
authors.initials: W
authors.lastname: Yan
authors.forename: Xinbin
authors.initials: X
authors.lastname: Chen
authors.forename: Chu-xia
authors.initials: CX
authors.lastname: Deng
authors.forename: Qin
authors.initials: Q
authors.lastname: Wang
authors.forename: Kai
authors.initials: K
authors.lastname: Jiao
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017209:Apoptosis; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000096:biosynthesis / Q000235:genetics; D002455:Cell Division; D005260:Female; D005315:Fetal Diseases / Q000209:etiology; D005318:Fetal Heart / Q000254:growth & development*; D018507:Gene Expression Regulation, Developmental; D005804:Genes, Lethal; D016259:Genes, myc; D006321:Heart / Q000196:embryology*; D006333:Heart Failure / Q000196:embryology / Q000209:etiology / Q000473:pathology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009206:Myocardium / Q000378:metabolism* / Q000473:pathology; D032383:Myocytes, Cardiac / Q000473:pathology; D041681:NIH 3T3 Cells; D038081:Organogenesis; D011401:Promoter Regions, Genetic; D016271:Proto-Oncogene Proteins c-myc / Q000096:biosynthesis / Q000502:physiology; D017386:Sequence Homology, Amino Acid; D051901:Smad4 Protein / Q000172:deficiency / Q000235:genetics / Q000502:physiology*; D020825:T-Box Domain Proteins / Q000096:biosynthesis / Q000235:genetics; D016212:Transforming Growth Factor beta / Q000096:biosynthesis / Q000235:genetics
chemical_list: C525718:BMP2 protein, human; C119152:Bmp10 protein, mouse; C525719:Bmp2 protein, mouse; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D016271:Proto-Oncogene Proteins c-myc; D051901:Smad4 Protein; C490810:Smad4 protein, mouse; C097348:T-Box Domain Protein 2; D020825:T-Box Domain Proteins; D016212:Transforming Growth Factor beta