• JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
 
  Bookmark and Share
 
 
Master's Dissertation
DOI
10.11606/D.17.2018.tde-29032018-102631
Document
Author
Full name
André Cendon Silva
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
Ribeirão Preto, 2017
Supervisor
Committee
Ramos, Ricardo Guelerman Pinheiro (President)
Allodi, Silvana
Hartfelder, Klaus Hartmann
Yan, Chao Yun Irene
Title in Portuguese
Estudo funcional das moléculas do módulo de reconhecimento celular Irre durante o desenvolvimento do sistema nervoso embrionário de Drosophila melanogaster
Keywords in Portuguese
Drosophila melanogaster
IRM
Linha média
Sistema nervoso
Abstract in Portuguese
O Módulo de Reconhecimento de Celular de Irre (IRM) é uma pequena família de proteínas transmembranares envolvidas nos processos de adesão e reconhecimento celular. Para Drosophila melanogaster existem 4 proteínas bem caracterizadas desta família: Rough (Rst), Hibris (Hbs), Kin de Irre (Kirre) e Stick and Stones (Sns). Em estudos anteriores para elucidar o papel dessas moléculas no desenvolvimento embrionário de Drosophila, observou-se que a construção pCa18 3.1, que apenas codifica a porção extracelular de Roughest e está sob controle do promotor de choque térmico, quando superexpressa no inicio do desenvolvimento embrionário gera defeitos na formação do sistema nervoso (NS), que são observados apenas mais tarde. Observou-se, além disso, que, quando esta superexpressão é realizada emum background onde apenas 50% da proteína Hbs está presente, o fenótipo é reforçado, sugerindo um possível papel antagonista entre Rst e Hbs no desenvolvimento de SN. Com base nestes achados e na informação que Rst, Hbs e Kirre são encontrados no SN durante o desenvolvimento embrionário, este trabalho tem como objetivo continuar estudando o papel dos IRMs no desenvolvimento da SN. Para este propósito, além do já empregado promotor de choque térmico, usaremos o sistema de expressão binária Gal4 / UAS com drivers para SN e, assim, gerando, além de superexpressão, também o knockdown por RNAi de IRMs. As construções genéticas contendo Dicer, uma enzima envolvida no processamento de RNAi, serão empregadas para melhorar o knockdown. A quantificação das transcrições será realizada por PCR em tempo real. Para estudos de co-localização e análise de fenótipos, técnicas de citocinética e imunocitoquímica serão empregadas.
Title in English
Functional study of Irre Cell Recognition Module molecules in developing embryonic nervous system of Drosophila melanogaster
Keywords in English
Drosophila melanogaster
IRM
Midline
Nervous system
Abstract in English
The Irre Cell Recognition Module (IRM) complex is a small family of transmembrane proteins involved in cellular adhesion and recognition processes. For Drosophila melanogaster there are 4 well characterized proteins of this family: Roughest (Rst), Hibris (Hbs), Kin of Irre (Kirre) and Stick and Stones (Sns). In previous studies to elucidate the role of these molecules in Drosophila embryonic development, it was noted that the construction pCa18 3.1, which only encodes the extracellular portion of Roughest and is under control of the heat-shock promoter, when overexpressed in early development generate defects in the formation of the nervous system (NS), which are observed only later. It was observed, furthermore, that when this overexpression is carried out in a mutant with a deficiency, in which the coding sequence Hbs is not present, this phenotype is enhanced, suggesting a possible antagonist role between Rst and Hbs in the development of SN. Based on these findings and the information that Rst, Hbs and Kirre are found in the SN during embryonic development, this work aims to further study the role of IRMs in the development of SN. For this purpose, besides the already employed heat-shock promoter, we will use the binary expression system Gal4/UAS with drivers for SN and, thereby, generating, in addition to overexpression, also the knockdown by RNAi of IRMs. Genetic constructs containing Dicer, an enzyme involved in the processing of RNAi, will be employed to enhance the knockdown. The quantification of the transcripts will be carried out by Real Time PCR. For co-localization studies and analysis of phenotypes techniques of cytochemistry and immunocytochemistry will be employed.
 
WARNING - Viewing this document is conditioned on your acceptance of the following terms of use:
This document is only for private use for research and teaching activities. Reproduction for commercial use is forbidden. This rights cover the whole data about this document as well as its contents. Any uses or copies of this document in whole or in part must include the author's name.
ANDRECENDONSILVA.pdf (829.99 Kbytes)
Publishing Date
2018-07-25
 
WARNING: Learn what derived works are clicking here.
All rights of the thesis/dissertation are from the authors
CeTI-SC/STI
Digital Library of Theses and Dissertations of USP. Copyright © 2001-2020. All rights reserved.