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Doctoral Thesis
DOI
10.11606/T.42.2008.tde-11112008-154726
Document
Author
Full name
Roberta Mascioli Cravo
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2008
Supervisor
Committee
Xavier Neto, José (President)
Alvares, Lucia Elvira
Franchini, Kleber Gomes
Scheepmaker, Denise Selivon
Yan, Chao Yun Irene
Title in Portuguese
Controle da expressão do gene ALDH1A2 (RALDH2) durante o desenvolvimento: uma abordagem filogenética.
Keywords in Portuguese
Biologia
Biologia do desenvolvimento
Embriologia
Embriologia molecular
Abstract in Portuguese
O ácido retinóico (AR) é essencial para a embriogênese. A principal enzima sintetizadora de AR durante o desenvolvimento é a ALDH1A2 (RALDH2), uma retinaldeído desidrogenase que converte retinaldeído a AR. Para entendermos como o gene da aldh1a2 é regulado identificamos regiões evolutivamente conservadas (ECRs) em vertebrados e testamos seu potencial regulatório. Identificamos uma ECR localizada no intron1 da aldh1a2, conservada em anfíbios, aves e mamíferos que atua como um enhancer em estruturas derivadas de ectoderme, endoderme e mesoderme. Animais transgênicos transientes e permanentes mostram a ativação desse enhancer na região da placa do teto do tubo neural e epicárdio, local onde esse enhancer é ativado em células derivadas do órgão pro-epicárdico após o contato e/ou proximidade com células do miocárdio. A identificação de um enhancer conservado no gene da aldh1a2 suporta a idéia de que esse gene possui uma regulação modular e mostra que a abordagem evolutiva é uma eficiente ferramenta para a identificação de mecanismos de controle desse gene.
Title in English
Search for regulatory elements in the ALDH1A2 (RALDH2) gene during development: a philogenetic approach.
Keywords in English
Biology
Developmental biology
Embryology
Molecular embryology
Abstract in English
Retinoic acid (RA) is essential for embryogenesis. The key RA synthetic enzyme during early development is ALDH1A2 (RALDH2), a retinaldehyde dehydrogenase that converts retinaldehyde into RA. To understand how aldh1a2 is regulated we screened the gene for evolutionary conserved regions (ECRs) among vertebrates and assayed their regulatory potential. We describe an aldh1a2 intron 1 ECR (identified as RALDH2.2) that is conserved in amphibians, avians and humans and acts as an enhancer in derivatives of ectoderm, endoderm and mesoderm. Transient and stable transgenesis in mice reveal strong activity of the raldh2 intron 1 enhancer at the roof plate of the neural tube and at the growing epicardium. Transgenic mice indicate that the enhancer is activated in proepicardium-derived cells by contact and/or close proximity to the myocardium. The identification of an aldh1a2 conserved enhancer supports the idea of a modular regulation and shows that the evolutionary approach is an efficient tool to identify control mechanisms of the aldh1a2 gene.
 
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Publishing Date
2008-11-12
 
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