Digital expression profiling of the compartmentalized translatome of Purkinje neurons.
Digital expression profiling of the compartmentalized translatome of Purkinje neurons.
复制标题
Purkinje神经元的分室翻译组的数字表达分析。
DOI：
10.1101/gr.164095.113
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发表时间：
2014-08
期刊：
影响因子：
7
通讯作者：
Launey T
中科院分区：
文献类型：
--
作者：
Kratz A;Beguin P;Kaneko M;Chimura T;Suzuki AM;Matsunaga A;Kato S;Bertin N;Lassmann T;Vigot R;Carninci P;Plessy C;Launey T
哺乳动物大脑的复杂性在于它的神经元连接网络，但也包括每个神经元内信号通路、蛋白质相互作用和受调控的基因表达的分子网络。空间混合神经元的多样性和复杂性给单个神经元成分的识别和量化带来了严峻的挑战。为了应对这一挑战，我们提出了一种新的方法来研究特定神经元的特定亚细胞域中的核糖体相关转录组-翻译组，并将其应用于大鼠小脑中的浦肯野细胞(PC)。我们结合显微解剖、在非转基因动物中翻译核糖体亲和纯化(TRAP)和定量纳米笼测序来获得PC及其树突中与细胞质或粗面内质网(RER)相关的核糖体结合的RNA的快照。这使我们能够发现PC的新标记，确定基因的结构方面，找到迄今尚未确定的转录本，并对控制离子稳态和神经元电活动的膜蛋白的生物物理相关基因进行量化。
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.
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影响因子：
14.9
作者：
Dougherty JD;Schmidt EF;Nakajima M;Heintz N
通讯作者：
Heintz N
影响因子：
30.8
作者：
Aldinger, Kimberly A.;Lehmann, Ordan J.;Hudgins, Louanne;Chizhikov, Victor V.;Bassuk, Alexander G.;Ades, Lesley C.;Krantz, Ian D.;Dobyns, William B.;Millen, Kathleen J.
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作者：
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影响因子：
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作者：
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通讯作者：
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影响因子：
14.9
作者：
Flicek P;Ahmed I;Amode MR;Barrell D;Beal K;Brent S;Carvalho-Silva D;Clapham P;Coates G;Fairley S;Fitzgerald S;Gil L;García-Girón C;Gordon L;Hourlier T;Hunt S;Juettemann T;Kähäri AK;Keenan S;Komorowska M;Kulesha E;Longden I;Maurel T;McLaren WM;Muffato M;Nag R;Overduin B;Pignatelli M;Pritchard B;Pritchard E;Riat HS;Ritchie GR;Ruffier M;Schuster M;Sheppard D;Sobral D;Taylor K;Thormann A;Trevanion S;White S;Wilder SP;Aken BL;Birney E;Cunningham F;Dunham I;Harrow J;Herrero J;Hubbard TJ;Johnson N;Kinsella R;Parker A;Spudich G;Yates A;Zadissa A;Searle SM
通讯作者：
Searle SM