RESUMO
OBJECTIVES: PAX9 belongs to the Pax family of transcriptional factor genes. This gene is expressed in embryonic tissues such as somites, pharyngeal pouch endoderm, distal limb buds and neural crest-derived mesenchyme. Polymorphisms in the upstream promoter region of the human PAX9 have been associated with human non-syndromic tooth agenesis. In the present study, we verified the in vitro mRNA expression of this gene and the luciferase activity of two constructs containing promoter sequences of the PAX9 gene. MATERIAL AND METHODS: Embryonic tissues were obtained from digits, face, and midbrain/hindbrain regions. Fragments containing PAX9 promoter sequences were cloned into reporter plasmids and were transfected into the different cell cultures. mRNA were extracted from primary cell cultures. RESULTS: The semi-quantitative RT-PCR results showed that in vitro E13.5 limb bud and CNS cells express PAX9, but cells derived from the facial region do not. Moreover, the luciferase assay showed that protein activity of the constructed vector was weaker than pgl3 -basic alone. CONCLUSIONS: The present results suggest that the promoter sequences analyzed are not sufficient to drive PAX9 gene transcription.
Assuntos
Anodontia/genética , Perfilação da Expressão Gênica , Luciferases/análise , Fator de Transcrição PAX9/genética , Transcrição Gênica , Animais , Células Cultivadas , Humanos , Luciferases/genética , Fator de Transcrição PAX9/metabolismo , Plasmídeos/genética , Regiões Promotoras Genéticas , RNA Mensageiro , Ratos , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
OBJECTIVES: PAX9 belongs to the Pax family of transcriptional factor genes. This gene is expressed in embryonic tissues such as somites, pharyngeal pouch endoderm, distal limb buds and neural crest-derived mesenchyme. Polymorphisms in the upstream promoter region of the human PAX9 have been associated with human non-syndromic tooth agenesis. In the present study, we verified the in vitro mRNA expression of this gene and the luciferase activity of two constructs containing promoter sequences of the PAX9 gene. MATERIAL AND METHODS: Embryonic tissues were obtained from digits, face, and midbrain/hindbrain regions. Fragments containing PAX9 promoter sequences were cloned into reporter plasmids and were transfected into the different cell cultures. mRNA were extracted from primary cell cultures. RESULTS: The semi-quantitative RT-PCR results showed that in vitro E13.5 limb bud and CNS cells express PAX9, but cells derived from the facial region do not. Moreover, the luciferase assay showed that protein activity of the constructed vector was weaker than pgl3 -basic alone. CONCLUSIONS: The present results suggest that the promoter sequences analyzed are not sufficient to drive PAX9 gene transcription.
Assuntos
Animais , Humanos , Ratos , Anodontia/genética , Perfilação da Expressão Gênica , Luciferases/análise , Fator de Transcrição PAX9/genética , Transcrição Gênica , Células Cultivadas , Luciferases/genética , Fator de Transcrição PAX9/metabolismo , Regiões Promotoras Genéticas , Plasmídeos/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , RNA MensageiroRESUMO
Qual profissional que no dia-a-dia de clínica não se confrontou com um paciente com um extenso tratamento odontológico a ser realizado e ao perguntar o porquê de não procurar os serviços de um profissional, ouve-se não uma queixa relacionada aos problemas financeiros, mas uma queixa sobre o profundo medo que é experimentado ao adentrar no consultório odontológico. Nem sempre as técnicas de manejo comportamental conseguem minimizar a sensação de temor perante o cirurgião-dentista, especialmente quando o paciente é uma criança ou adulto que apresenta traumas relacionados a um tratamento odontológico e/ou médico. Diante de tal fato, encontra-se na utilização do óxido nitros%xigênio um aliado. É feita uma revisão da literatura sobre com o objetivo de familiarizar o cirurgião-dentista sobre a técnica de controle da ansiedade...
