RESUMO
UNLABELLED: Nuclear factor kappaB (NF-kappaB) is a pleiotropic transcription factor implicated in the regulation of diverse morphologic cardiac alterations, for which the p50 and p65 subunits form the most prevalent dimeric form in the heart. NF-kappaB is inactivated by proteins of the IkappaB family, which trap it in the cytoplasm. It is not known whether NF-kappaB influences cardiac development. OBJECTIVE: Here we investigated the role of NF-kappaB in regulating transcription in chicken heart morphogenesis. Specifically, we tested whether NF-kappaB activation is required for normal formation of the outflow tract (OFT) during a critical stage of heart development. METHODS AND RESULTS: We designed a reporter vector with kappaB binding sites for Rel family members in the promoter, upstream from the cDNA of Green Fluorescent Protein (GFP). This construct was injected directly into the developing heart of chicken embryos. NF-kappaB activation was subsequently inhibited by administration of the specific pharmacological agent Bay 11-7085. We found that forced NF-kappaB expression was associated with multiple congenital cardiac alterations of the OFT (mainly IVC, DORV and great arteries stenosis). CONCLUSION: These findings indicate that blockade of NF-kappaB induces apoptosis and is an important factor in the development of OFT during cardiogenesis. However, it remains unknown which members of the Rel family are relevant in this process.
Assuntos
Coração/efeitos dos fármacos , NF-kappa B/metabolismo , Nitrilas/farmacologia , Transdução de Sinais/efeitos dos fármacos , Sulfonas/farmacologia , Animais , Apoptose/efeitos dos fármacos , Linhagem Celular , Células Cultivadas , Embrião de Galinha , Meios de Cultura Livres de Soro/farmacologia , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Coração/embriologia , Coração/fisiologia , Humanos , Modelos Biológicos , Morfogênese/efeitos dos fármacos , Miócitos Cardíacos/citologia , Miócitos Cardíacos/efeitos dos fármacos , Miócitos Cardíacos/metabolismo , NF-kappa B/antagonistas & inibidores , NF-kappa B/genética , Teratogênicos/farmacologia , Transfecção , Fator de Necrose Tumoral alfa/farmacologiaRESUMO
A Mexican population sample was obtained from the central region of the country, including five states. Two hundred and eleven individuals were PCR-typed for 15 STR loci with the AmpFiota STR Identifiler PCR amplification kit (Applied Biosystems). The following autosomal markers were analyzed: D8S1179, D21S11, D7S820, CSF1PO, D19S433, vWA, TPOX, D18S51, D3S1358, TH01, D13S317, D16S539, D2S1338, D5S818, FGA and Amelogenin. Allele frequencies for each STR were estimated and compared to previous reports. Genotype distribution by locus and by two loci combination was in agreement with Hardy-Weinberg expectations for all 15 STRs. This STR system in Mexican-mestizos presented a combined probability of exclusion (PE) and discrimination (PD) longer than 99.999%.
