Cloning and expression analysis of the transcription factor Scratch2 during the chicken early embryogenesis. / Clonagem e análise da expressão do fator de transcrição Scratch2 durante a embriogênese inicial de galinha.

AUTOR(ES)

Felipe Monteleone Vieceli

DATA DE PUBLICAÇÃO

2009

RESUMO

In invertebrates, the Scratch (Scrt) genes encode transcription factors that promote neurogenesis during development. The Scrt function in vertebrates is unknown, but in mice Scrt1 and Scrt2 are specifically expressed in post-mitotic neurons in the embryo and in the adult central nervous system. In this work, we have cloned the coding sequence of chicken Scrt2 (cScrt2) and characterized its expression pattern in the embryo with quantitative PCR and in situ hybridization. The complete coding sequence was cloned in the expression vector pMES-GFP and the predicted translation product is a 276-aminoacids protein. The aminoacid sequence shares identities of 70% with rat Scrt2 and 58% with zebrafish Scrt. cScrt2 transcripts are firstly detected in the periphery of the neural tube in the hindbrain by HH 15 and in the spinal cord by HH 17, coinciding with the places where some of the first neurons differentiate during embryogenesis. Between HH 19-23, the expression in the motor domain of the spinal cord is progressively concentrated in the interface between the ventricular and mantle zones. Furthermore, cScrt2 expression is also observed in the dorsal root ganglia after HH22-23, particularly in the dorsomedial domain. The expression pattern of cScrt2 in the neural tube is complementary to that of Notch1, which is expressed in neural stem cells, and SCG10, a marker for differentiated neurons. Our results suggest that during embryogenesis cScrt2 is specifically expressed in post-mitotic undifferentiated neurons. The construction pMES-GFP(cScrt2) makes possible future direct functional analysis by genetic interference in the chick embryo, which will be of great value for better understanding the Scrt genes function in vertebrates.

ASSUNTO(S)

cell diferentiation clonagem molecular hibridização molecular embryology medula espinhal hybridization fatores de transcrição spinal cord transcription factors embriologia molecular diferenciação celular molecular cloning

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