RT Journal
A1 Kratz, Anton
A1 Beguin, Pascal
A1 Kaneko, Megumi
A1 Chimura, Takahiko
A1 Suzuki, Ana Maria
A1 Matsunaga, Atsuko
A1 Kato, Sachi
A1 Bertin, Nicolas
A1 Lassmann, Timo
A1 Vigot, Réjan
A1 Carninci, Piero
A1 Plessy, Charles
A1 Launey, Thomas
T1 Digital expression profiling of the compartmentalized translatome of Purkinje neurons
JF Genome Research 
JO Genome Research 
YR 2014 
FD August 01 
VO 24 
IS 8 
SP 1396 
OP 1410 
DO 10.1101/gr.164095.113 
UL http://genome.cshlp.org/content/24/8/1396.abstract 
AB 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 pose 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 subcellular domains of specific neurons, and apply it to the Purkinje cells (PCs) in the rat cerebellum. We combined microdissection, translating ribosome affinity purification (TRAP) in nontransgenic 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.