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Doctoral Thesis
DOI
https://doi.org/10.11606/T.42.2014.tde-13082014-151401
Document
Author
Full name
Jamile Calil Silveira
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2014
Supervisor
Committee
Nunes, Maria Tereza (President)
Carvalho, Denise Pires de
Chaves, Maria Luiza Morais Barreto de
Geraldo, Murilo Vieira
Rubio, Ileana Gabriela Sanchez de
Title in Portuguese
Efeito da administração aguda de iodo na expressão gênica e atividade do promotor da pendrina: estudos em ratos e em células PCCI3.
Keywords in Portuguese
Efeito Wolff-Chaikoff
Escape
Expressão gênica
Iodo
Pendrina
Tiróide
Abstract in Portuguese
Pendrina é um transportador de iodo na tiróide, codificada pelo gene PDS.Sabendo-se que o iodo é fundamental para a síntese de T3 e T4 foi objetivo avaliar o efeito do iodo na expressão gênica, atividade e promotor da pendrina. Foram utilizados ratos Wistar, que receberam injeção de salina ou 2mg de NaI. Após 30, 1, 24 e 48h as tiróides foram removidas para análise do mRNA e proteína, por PCR em Tempo Real e Western Blot. Ainda, utilizou-se celulas PCCl3, que foram tratadas ou não com 10-3M de NaI. Após 30, 1, 24 e 48h o RNA/proteína foram isolados.Determinou-se o efluxo pela medição do125I presente no meio de cultura. Para analisar a atividade do promotor, ele foi inserido no vetor pGL3-básico.Nossos resultados mostram que houve aumento do mRNA da pendrina em resposta ao iodo em todos os tempos estudados, porém não houve aumento da atividade promotora.A expressão da proteína pendrina, bem como taxa do efluxo de iodeto, aumentou nos tempos mais longos de tratamento.Esses dados apontam um importante papel de pendrina para o fenômeno de autoregulação da tiróide.
Title in English
Effect of acute administration of iodine on pendrin gene expression and the activity of pendrin promoter: studies in rats and PCCl3 cells.
Keywords in English
Escape
Gene expression
Iodine
Pendrin
Thyroid
Wolff-Chaikoff effect
Abstract in English
Pendrin is an iodide transporter in thyroid, encoded by the PDS gene. Since iodide is essential for the synthesis of T3 and T4, this study aimed to evaluate the iodine effect on pendrin gene expression and on the activity of its promoter. Male rats received an injection of saline or 2mg of NaI. After 30, 1, 24 and 48h the thyroids were removed for mRNA and protein analysis by Real Time PCR and Western Blot. Moreover, PCCl3 cells were treated or not with 10-3M NaI. After 30, 1, 24 and 48h the RNA/protein were isolated. The efflux was determined by measure of 125I in the culture medium. For the promoter analysis, it was inserted in pGL3-basic plasmid. The results indicated that the pendrin mRNA amount increased after iodide treatment. However, the promoter activity was unchanged. The protein expression and iodide efflux increased only after 24h of treatment. These data suggest an important role of pendrin for the autoregulation of the thyroid.
 
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Publishing Date
2014-08-14
 
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