Análise do transcriptoma durante a ontogenia do timo. / Analysis of the transcriptoma during the antogenia of the thymus.
AUTOR(ES)
Danielle Aparecida Rosa de Magalhães
DATA DE PUBLICAÇÃO
2007
RESUMO
The thymus is a complex organ structured by a stroma, which is formed mainly by cortical epithelial cells (cTECs) and medullary epithelial cells (mTECs) besides of other cell types such as dendritic cells (DC), macrophages, B lymphocytes and fibroblasts. Moreover, the T cell precursors arising from the bone marrow reach the thymus (thymocytes) maturing in T lymphocytes, which migrate to the periphery. Thus, the thymus is the place of very important events during the maturation of the immune system, including the control of their own homeostasis. In the present study, we search to picture the main characteristics of the thymus by means of the large scale gene expression analysis that is, describing part of their transcriptome. We made use of the cDNA microarray technology in two versions. In the first one we used cDNA microarrays constructed on glass slides and fluorescent probes labeled with the fluorochromes Cy3 or Cy5 and in the second version we used cDNA microarrays on nylon membranes and radioactive probes labeled with 33P isotope. To data analysis we used dedicated bioinformatics programs, such as SAM (significance analysis of microarrays) and Cluster-Tree View. Three sets of results were possible. In the first set we observed the occurrence of the promiscuous gene expression (PGE) of parenchymal tissue/organ specific antigens (TSAs), demarking their temporal emergence during the murine thymus ontogeny, which is influenced by the genetic background of the inbred strains studied. The occurrence of PGE in the thymus is associated to the molecular-genetics basis of the immune tolerance of T cells, contributing with the prevention of autoimmunity. The second set of results consisted in the analysis of gene expression of thymus from knockout mice (KO) involving genes important for T cell maturation, such as TCR?, LAT, Relb, RAG-1 and CD3?, allowing the observation of its effects on the transcription regulation in this organ. Finally, the third set consisted in the definition of the virtual molecular dissection of the thymus. By means of particular gene expression profiling featured by each main cell type populating the thymus, it was possible to dissect this organ using the cDNA microarray technology.
ASSUNTO(S)
thymus expressão gênica promíscua (pge) promiscuous gênica expression (pge) timo microarranjos de cdna microarranjos of cdna
