O papel modulador do gene AIRE - capes

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T cell signaling gene networks during thymus developmentGenes coding for T cell signaling proteins establishtranscriptional regulatory networks during thymus ontogenyCláudia Macedo 1 + , Danielle A. Magalhães 1 + , Monique Tonani 1 ,Márcia C. Marques 1 , Cristina M. Junta 1 , Geraldo A. S. Passos 1,2 *1) Molecular Immunogenetics Group, Department of Genetics, Facultyof Medicine, University of São Paulo (USP), 14040-900, RibeirãoPreto, SP, Brazil.2) Discipline of Genetics, Department of Morphology (DMEF), Facultyof Dentistry, USP, 14040-904, Ribeirão Preto, SP, Brazil.+ These authors contributed equally to this study* Send correspondence to:Dr. Geraldo A. S. PassosMolecular Immunogenetics GroupDepartment of GeneticsFaculty of Medicine of Ribeirão Preto, USP14040-900 Ribeirão Preto, SP, Brazil.Tel +55 16 3602 3030 Fax +55 16 3633 0069E-mail passos@rge.fmrp.usp.br235
AbstractThe definition of gene expression profiling by cDNA microarrays during murinethymus ontogeny has contributed to dissecting the large-scale moleculargenetics of T cell maturation. Gene profiling, although useful for characterizingthe thymus developmental phases and identifying the differentially expressedgenes, does not permit the determination of possible interactions betweengenes. In order to reconstruct genetic interactions during a murine thymusperiod of development, which is important for thymocyte differentiation (14-17days of gestation), a pair of microarrays was used containing a total of 1,576cDNA sequences derived from the IMAGE MTB library, whose data wereanalyzed using the GeneNetwork program. Characteristic interactions involvinggenes were observed during thymus ontogeny, which were previously identifiedas differentially expressed and once included in a network, showed theirrelationships with several other genes. The present method provided thedetection of gene nodes implicated in the calcium signaling pathway, such asPrr2 and Stxbp3, and in protein transport towards the cell membrane, such asGosr2. The results demonstrate the feasibility of reconstructing networks basedon cDNA microarray gene expression determinations, contributing to a clearerunderstanding of the complex interactions between genes involved inthymus/thymocyte development.Keywords Thymus development, gene expression, microarray, gene networks236
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Universidade de São PauloFaculdade
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Macedo, ClaudiaO papel modulador do
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Dedico Especialmente este TrabalhoM
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AgradecimentosAgradeço do fundo do
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ÍndiceRESUMO .....................
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INDICE DE FIGURASFigura 1. Desenvol
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Promiscuous gene expression in medu
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INTRODUÇÃO1. INTRODUÇÃO1.1. A m
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INTRODUÇÃORecentemente surgiram e
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INTRODUÇÃOmigram para a região s
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INTRODUÇÃOCD4 + CD8 + , e Ccl21 (
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INTRODUÇÃOos timócitos duplo-pos
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INTRODUÇÃOgrande diversidade nas
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INTRODUÇÃOAs células mTECs são
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INTRODUÇÃODerbinski (2004). Os ti
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INTRODUÇÃOinvertebrados multicelu
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INTRODUÇÃOCoraçãoLínguaEstôma
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INTRODUÇÃO1.3. O papel do gene Au
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INTRODUÇÃOdefinido pela atividade
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INTRODUÇÃOpara abordarmos nossa p
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INTRODUÇÃOesses modelos levam em
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Hipóteses
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Objetivos
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Material e Métodos
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MATERIAL E MÉTODOSFigura 6. Padrã
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MATERIAL E MÉTODOScom estes RNAs f
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MATERIAL E MÉTODOSdos complexos de
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MATERIAL E MÉTODOSAo RNA total rec
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4.5. PCRs semi-quantitativas para o
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MATERIAL E MÉTODOSminutos a 4 °C.
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MATERIAL E MÉTODOS(Merck), soluç
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MATERIAL E MÉTODOSEnsaios de co-hi
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MATERIAL E MÉTODOSRNA TotalANELAME
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MATERIAL E MÉTODOSa) Preparação
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MATERIAL E MÉTODOScentrifugação
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MATERIAL E MÉTODOScanais (Cy3 e Cy
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MATERIAL E MÉTODOSSpotfinderQuanti
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MATERIAL E MÉTODOSOnde Xp(i) e Xc(
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MATERIAL E MÉTODOSO programa SAM p
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MATERIAL E MÉTODOSextrair automati
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DELINEAMENTO EXPERIMENTAL5. DELINEA
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RESULTADOS6. RESULTADOS6.1. Inibiç
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RESULTADOSPCR semiquantitativa gene
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RESULTADOS6.3. RT-PCRs semi-quantit
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RESULTADOS11.00%6,50%AdequadaNão A
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RESULTADOS6.6. Quantificação e no
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RESULTADOSPara um melhor entendimen
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RESULTADOSFigura 25. Agrupamento hi
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RESULTADOSFigura 26. Gráfico ilust
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RESULTADOS6.8. Influência do silen
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Discussão
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DISCUSSÃOA tolerância do repertó
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DISCUSSÃOCom relação à técnica
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DISCUSSÃOgene AIRE (do inglês aut
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DISCUSSÃOdesse modo a tolerância
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DISCUSSÃOPML (leucemia promielocit
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DISCUSSÃOBoehm et al. (2003) afirm
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DISCUSSÃOprograma Genenetwork (Wu
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Conclusões
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Referências Bibliográficas
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REFERÊNCIAS BIBLIOGRÁFICASBarthlo
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REFERÊNCIAS BIBLIOGRÁFICASDissert
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REFERÊNCIAS BIBLIOGRÁFICASGotter
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REFERÊNCIAS BIBLIOGRÁFICASMatsumo
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REFERÊNCIAS BIBLIOGRÁFICASby mito
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REFERÊNCIAS BIBLIOGRÁFICASRossi S
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REFERÊNCIAS BIBLIOGRÁFICASof thym
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Anexo I123
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ANEXO IEST IMAGE:582370 Expressed s
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ANEXO IDhps IMAGE:583332 Deoxyhypus
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ANEXO IMapkapk2 IMAGE:572979 MAP ki
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ANEXO IGm608 IMAGE:583350 Gene mode
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ANEXO ICcdc72 IMAGE:640628 Coiled-c
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ANEXO ITranscribed locusTranscribed
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ANEXO ITranscribed locusTranscribed
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ANEXO Irepair deficiency, complemen
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ANEXO Imember 1EST IMAGE:583824 Exp
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ANEXO ITranscribed locus IMAGE:5830
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ANEXO ITfdp1 IMAGE:640119 Transcrib
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ANEXO I2410022M11Rik IMAGE:640727 R
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ANEXO Icontaining 1Kif14 IMAGE:5836
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ANEXO Imember 3In multiple clusters
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ANEXO IAnkrd10 IMAGE:583740 Ankyrin
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ANEXO IVcam1 IMAGE:576563 Vascular
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ANEXO ITegt IMAGE:639877 Testis enh
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ANEXO I3Ehmt2Euchromatic histone ly
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ANEXO I20 homolog (yeast)Ube1c IMAG
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ANEXO IRasa3 IMAGE:582174 RAS p21 p
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ANEXO I4930566A11Rik IMAGE:583433 R
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Page 200 and 201: Anexo II
Page 202 and 203: ANEXO II1500002B03RikIMAGE:1330385
Page 204 and 205: ANEXO IIEST IMAGE:640173 Expressed
Page 206 and 207: ANEXO IICnot10 IMAGE:576406 CCR4-NO
Page 208 and 209: ANEXO IIEST IMAGE:576142 Expressed
Page 210 and 211: Anexo III
Page 212 and 213: ANEXO III197Ifit1RelaIMAGE:575632Gr
Page 214 and 215: ANEXO III199Nde1Nol5aIMAGE:640341Rp
Page 216 and 217: Manuscrito da Tese
Page 218 and 219: MANUSCRITOAbstractThe expression of
Page 220 and 221: MANUSCRITOdominated the scenario in
Page 222 and 223: MANUSCRITOserum autoantibodies, all
Page 224 and 225: MANUSCRITOwere transfected with 10
Page 226 and 227: MANUSCRITOwere analyzed using the M
Page 228 and 229: MANUSCRITOpurified from ex vivo mou
Page 230 and 231: MANUSCRITOinformation theory as ARA
Page 232 and 233: MANUSCRITOKyewski B, Derbinski J, G
Page 234 and 235: MANUSCRITOBruno R, Sabater L, Tolos
Page 236 and 237: MANUSCRITOFigure 3. The gene expres
Page 238 and 239: MANUSCRITOABFigure 5. Genetics netw
Page 240 and 241: SÚMULACURRICULUM VITAE (Janeiro 20
Page 242 and 243: ESTÁGIO NO EXTERIORSÚMULA• Labo
Page 244 and 245: SÚMULAtranscript finishing initiat
Page 246 and 247: SÚMULAlinhagens isogênicas de cam
Page 248 and 249: SÚMULABOLSAS DE ESTUDO• Bolsa de
Page 252 and 253: IntroductionThe thymus is an import
Page 254 and 255: ontogeny using such technology has
Page 256 and 257: laboratory using a Generation III A
Page 258 and 259: cDNA microarray data analysisStatis
Page 260 and 261: Gene networksAs previously mentione
Page 262 and 263: cells, including human herpesvirus-
Page 264 and 265: gene was mostly indirectly positive
Page 266 and 267: lymphocytes and finally associates
Page 268 and 269: 4) Finally, Prrg2 was also identifi
Page 270 and 271: 14. Kishimoto H, Sprent J (2000) Th
Page 272 and 273: proliferation of a rat astrocyte ce
Page 274 and 275: IMMUNOLOGYORIGINAL ARTICLEOnset of
Page 276 and 277: Promiscuous gene expression in muri
Page 282 and 283: R.S. Cardoso et al. / Molecular Imm
Page 290 and 291: Molecular Immunology 42 (2005) 1043
Page 292 and 293: D.A.R. Magalhães et al. / Molecula
Page 294 and 295: D.A.R. Magalhães et al. / Molecula
Page 296 and 297: Molecular and Cellular Biochemistry
Page 298 and 299: 225Table 1. Target cDNA sequences a
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227coincided (CBA/J) with expressio
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