||In vivo analysis of processing bodies using transgenic mice expressing GFP-Dcp1a
塚本, 智史 ,
原, 太一 ,
佐藤, 健 ,
南, 直治郎 ,
鬼頭, 靖司小久保, 年章
The control of mRNA translation and degradation is important in the regulation of eukaryotic gene expression. Processing bodies (P-bodies) are cytoplasmic foci where mRNA decapping occurs and are therefore closely linked to mRNA degradation, storage, and translation. Although it is known that the number and size of P-bodies change in response to cellular stress including cell cycle arrest,DNA damage and senescence, their physiological functions are largely nknown. In this study,we generated transgenic mice that ubiquitously expressed GFP fused with the decapping enzyme Dcp1a under the control of the CAG promoter. GFP-Dcp1a was expressed in almost all tissues examined, with higher levels detected in heart, muscle, and testis. Immunofluorescence analysisin frozen tissue sections revealed the co-localization of GFP-Dcp1a and EDC4, an endogenous P-body marker. We also confirmed the loss of P-bodies by cycloheximide treatment of embryonic fibroblasts isolated from the transgenic mice. We further analyzed the changes in P-bodies during heart development and found that their numbers dramatically increased after birth and then de-creased in a time-dependent manner. Taken together, our results show that this transgenic mouse provides a tool for investigating the physiological functions of mammalian P-bodies in a wide range of research field.
The 15th International Congress of Radiation Research (ICRR 2015)