Agonist-induced functional analysis and cell sorting associated with single-cell transcriptomics characterizes cell subtypes in normal and pathological brain.
Agonist-induced functional analysis and cell sorting associated with single-cell transcriptomics characterizes cell subtypes in normal and pathological brain.
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DOI：
10.1101/gr.262717.120
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发表时间：
2020-11
期刊：
影响因子：
7
通讯作者：
Maurin T
中科院分区：
文献类型：
--
作者：
Castagnola S;Cazareth J;Lebrigand K;Jarjat M;Magnone V;Delhaye S;Brau F;Bardoni B;Maurin T
为了更好地了解细胞与其环境之间的动态相互作用，我们开发了激动剂诱导的功能分析和细胞分选（aiFACS）技术，该技术允许根据细胞对刺激的即时和个体反应同时实时记录和分选细胞。通过调节 aiFACS 选择参数、测试不同的发育时间、使用各种刺激以及增加读数分析，可以分析任何正常或病理组织的细胞群。 aiFACS 与单细胞转录组学的结合允许基于细胞的特定药理反应构建功能组织制图。作为概念验证，我们在分离的小鼠大脑（一种高度异质的组织）上使用 aiFACS，通过用 KCl 或 AMPA（谷氨酸受体激动剂）刺激来富集中间神经元，然后根据钙水平进行分类。 AMPA 刺激后，这些 aiFACS 选择的中间神经元的单细胞转录组学产生了九簇分类。此外，我们对源自 Fmr1-KO 小鼠大脑的中间神经元使用了 aiFACS，Fmr1-KO 小鼠是一种脆性 X 综合征的啮齿动物模型。我们发现，与野生型细胞相比，这些中间神经元对 AMPA 表现出普遍的缺陷反应，在一个特定的出生后发育时间影响所有分析的细胞簇。
To gain better insight into the dynamic interaction between cells and their environment, we developed the agonist-induced functional analysis and cell sorting (aiFACS) technique, which allows the simultaneous recording and sorting of cells in real-time according to their immediate and individual response to a stimulus. By modulating the aiFACS selection parameters, testing different developmental times, using various stimuli, and multiplying the analysis of readouts, it is possible to analyze cell populations of any normal or pathological tissue. The association of aiFACS with single-cell transcriptomics allows the construction of functional tissue cartography based on specific pharmacological responses of cells. As a proof of concept, we used aiFACS on the dissociated mouse brain, a highly heterogeneous tissue, enriching it in interneurons by stimulation with KCl or with AMPA, an agonist of the glutamate receptors, followed by sorting based on calcium levels. After AMPA stimulus, single-cell transcriptomics of these aiFACS-selected interneurons resulted in a nine-cluster classification. Furthermore, we used aiFACS on interneurons derived from the brain of the Fmr1-KO mouse, a rodent model of fragile X syndrome. We showed that these interneurons manifest a generalized defective response to AMPA compared with wild-type cells, affecting all the analyzed cell clusters at one specific postnatal developmental time.
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影响因子：
46.9
作者：
Fuzik J;Zeisel A;Máté Z;Calvigioni D;Yanagawa Y;Szabó G;Linnarsson S;Harkany T
通讯作者：
Harkany T
影响因子：
4.3
作者：
Abekhoukh S;Sahin HB;Grossi M;Zongaro S;Maurin T;Madrigal I;Kazue-Sugioka D;Raas-Rothschild A;Doulazmi M;Carrera P;Stachon A;Scherer S;Drula Do Nascimento MR;Trembleau A;Arroyo I;Szatmari P;Smith IM;Milà M;Smith AC;Giangrande A;Caillé I;Bardoni B
通讯作者：
Bardoni B
影响因子：
3.7
作者：
Zwartz, Gordon J.;Chigaev, Alexandre;Foutz, Terry D.;Edwards, Bruce;Sklar, Larry A.
通讯作者：
Sklar, Larry A.
影响因子：
56.9
作者：
FULWYLER, MJ
通讯作者：
FULWYLER, MJ
影响因子：
46.9
作者：
Cadwell CR;Palasantza A;Jiang X;Berens P;Deng Q;Yilmaz M;Reimer J;Shen S;Bethge M;Tolias KF;Sandberg R;Tolias AS
通讯作者：
Tolias AS