Establishment of a model of murine odontoblasts underexpressing Pkd1 using shRNA

Authors

  • B Thivichon-Prince
  • N Labert
  • M L Couble
  • F Bleicher

Keywords:

odontoblast, Pkd1 down expression, mechanotransduction

Abstract

We have previously shown that PKD1, the gene encoding Polycystin-1 (or TRPP1) is expressed in human odontoblasts and that this protein is localized at the primary cilium of the cell. Nevertheless, its function remain unclear in this cell even if studies on osteoblasts, osteocytes and chondrocytes give TRPP1 as a promising candidate for mechanotransduction in response to mechanical stress. Consequently, to evaluate the role of TRPP1 in this transduction process, we needed first to generate an in vitro murine model down expressing Pkd1. Using lentivirus-mediated shRNA technology, we obtained a 60% suppression of Pkd1 mRNA expression in transfected MO6-G3 cells associated with a decrease of cell proliferation. Thus, establishment of this murine odontoblast model underexpressing Pkd1 associated with applied mechanical forces (compression or shear stress) will allow us to go further in the determination of TRPP1 involvement in odontoblasts mechanotransduction.

Nous avons montré précédemment que PKD1, le gène codant pour la polycystine 1 ou TRPP1, est exprimé dans les cultures d’odontoblastes humains avec une localisation préférentielle de la protéine au niveau des cils primaires. Cependant, la fonction de TRPP1 reste à ce jour inconnue malgré un rôle potentiel de mécanotransducteur mis en évidence dans les ostéoblastes, ostéocytes et chondrocytes. Pour évaluer le rôle de cette protéine dans les odontoblastes nous avons choisi de mettre au point un modèle cellulaire sous-exprimant Pkd1. Ainsi, à l’aide de shRNA, nous avons obtenu une lignée cellulaire de souris (MO6-G3) sous-exprimant de façon stable Pkd1 (60% de sous-expression), et présentant une diminution de la prolifération cellulaire. Ce nouveau modèle cellulaire associé à l’application de forces mécaniques (compression ou étirement) devrait nous permettre d’évaluer l’implication de TRPP1 dans les processus de mécanotransduction des odontoblastes.

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Issue

Vol. 52 No. 1 (2013)

Section

Original Research Articles

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Further, I certify that:

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5. The copyright of the article is transferred from the authors to the Bulletin du Groupement International pour la Recherche Scientifique en Stomatologie et Odontologie upon acceptance of the manuscript.