Laser capture microdissection enables cellular and molecular studies of tooth root development / 国际口腔科学杂志·英文版
International Journal of Oral Science
;
(4): 7-13, 2012.
Artigo
em Inglês
| WPRIM
| ID: wpr-269645
ABSTRACT
Epithelial-mesenchymal interactions (EMIs) are critical for tooth development. Molecular mechanisms mediating these interactions in root formation is not well understood. Laser capture microdissection (LCM) and subsequent microarray analyses enable large scale in situ molecular and cellular studies of root formation but to date have been hindered by technical challenges of gaining intact histological sections of non-decalcified mineralized teeth or jaws with well-preserved RNA. Here,we describe a new method to overcome this obstacle that permits LCM of dental epithelia,adjacent mesenchyme,odontoblasts and cementoblasts from mouse incisors and molars during root development. Using this method,we obtained RNA samples of high quality and successfully performed microarray analyses. Robust differences in gene expression,as well as genes not previously associated with root formation,were identified. Comparison of gene expression data from microarray with real-time reverse transcriptase polymerase chain reaction (RT-PCR) supported our findings. These genes include known markers of dental epithelia,mesenchyme,cementoblasts and odontoblasts,as well as novel genes such as those in the fibulin family. In conclusion,our new approach in tissue preparation enables LCM collection of intact cells with well-preserved RNA allowing subsequent gene expression analyses using microarray and RT-PCR to define key regulators of tooth root development.
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Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Fisiologia
/
Germe de Dente
/
Raiz Dentária
/
Regulação da Expressão Gênica no Desenvolvimento
/
Reação em Cadeia da Polimerase Via Transcriptase Reversa
/
Análise de Sequência com Séries de Oligonucleotídeos
/
Biologia Celular
/
Cemento Dentário
/
Transição Epitelial-Mesenquimal
/
Microdissecção e Captura a Laser
Tipo de estudo:
Estudo prognóstico
Limite:
Animais
Idioma:
Inglês
Revista:
International Journal of Oral Science
Ano de publicação:
2012
Tipo de documento:
Artigo
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