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Gene Expression and Translational Regulation

開催日 2014/9/11
時間 18:15 - 19:00
会場 Room J(313+314)
Chairperson(s) 奥野 浩行 / Hiroyuki Okuno (京都大学大学院医学研究科 メディカルイノベーションセンター / Medical Innovation Center, Kyoto University Graduate School of Medicine, Japan)
重本 隆一 / Ryuichi Shigemoto (IST Austria, Austria)

Purkinje neuron translatome at subcellular resolution

  • O1-J-6-1
  • Thomas Launey:1 Anton Kratz:2 Pascal Beguin:1 Megumi Kaneko:1 Takahiko Chimura:1 Ana Maria Suzuki:2 Atsuko Matsunaga:1 Sachi Kato:2 Nicolas Bertin:2 Timo Lassmann:2 Réjan Vigot:1 Piero Carninci:1 Charles Plessy:2 
  • 1:Launey Research Unit, Brain Sci.Inst., RIKEN, Wako-shi, Japan 2:RIKEN Center for Life Science Technologies, Division of Genomic Technologies, Yokohama, Japan 

Underlying the complexity of the mammalian brain is its network of neuronal connections, but also the molecular networks of signaling pathways, protein interactions and regulated gene expression within each individual neuron. The diversity and complexity of the spatially intermingled neurons poses a serious challenge to the identification and quantification of single neuron components. To address this challenge, we present a novel approach for the study of the ribosome-associated transcriptome - the translatome - from selected sub-cellular domains of specific neurons, and apply it to the Purkinje cells (PC) in the rat cerebellum.
We combined microdissection, translating ribosome affinity purification (TRAP) in non-transgenic animals and quantitative nanoCAGE sequencing to obtain a snapshot of RNAs bound to cytoplasmic or rough endoplasmic reticulum (rER)-associated ribosomes, in the PC and its dendrites. This allowed us to discover novel markers of PCs, to determine structural aspects of genes, to find hitherto uncharacterized transcripts, and to quantify biophysically relevant genes of membrane proteins controlling ion homeostasis and neuronal electrical activities.

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