A Molecular Calcium Integrator Reveals a Striatal Cell Type Driving Aversion.
A Molecular Calcium Integrator Reveals a Striatal Cell Type Driving Aversion.
复制标题
DOI：
10.1016/j.cell.2020.11.015
复制
发表时间：
2020-12-23
期刊：
影响因子：
64.5
通讯作者：
Ting, Alice Y.
中科院分区：
文献类型：
--
作者：
Kim, Christina K.;Sanchez, Mateo, I;Hoerbelt, Paul;Fenno, Lief E.;Malenka, Robert C.;Deisseroth, Karl;Ting, Alice Y.
在细胞的DNA或RNA中记录瞬时细胞事件的能力将使异质细胞群体的精确、大规模分析、选择和重编程成为可能。在这里，我们报告了一种分子技术，用于细胞内钙浓度（FLiCRE）活性相关增加的细胞的稳定遗传标记。我们使用FLiCRE转录标记小鼠大脑伏隔核在短暂的厌恶输入刺激期间激活的神经系统。通过单细胞RNA测序，我们检测了内源性转录组中的FLiCRE转录本，同时读出了细胞类型和钙活化历史。我们发现伏隔核中有一种细胞类型在远距离兴奋性投射的下游被激活。利用FLiCRE的模块化设计，我们在这种细胞类型中选择性地表达了光遗传通道，并表明直接招募这种遗传上不可接近的群体会引起行为厌恶。FLiCRE的特异性和分钟分辨率使激活细胞集合的分子信息表征，操作和重编程成为可能。Kim等人提出了flre (Fast Light and Calcium-Regulated Expression)，这是一种稳定的基因工程工具，可以在几分钟内对小鼠大脑中的细胞群进行活动依赖性标记。在这里，FLiCRE被用来识别和进一步表征伏隔核中参与驱动行为厌恶的瞬时激活神经元。
The ability to record transient cellular events in the DNA or RNA of cells would enable precise, large-scale analysis, selection, and reprogramming of heterogeneous cell populations. Here we report a molecular technology for stable genetic tagging of cells that exhibit activity-related increases in intracellular calcium concentration (FLiCRE). We used FLiCRE to transcriptionally label activated neural ensembles in the nucleus accumbens of the mouse brain during brief stimulation of aversive inputs. Using single-cell RNA sequencing, we detected FLiCRE transcripts among the endogenous transcriptome, providing simultaneous readout of both cell-type and calcium activation history. We identified a cell-type in the nucleus accumbens activated downstream of long-range excitatory projections. Taking advantage of FLiCRE’s modular design, we expressed an optogenetic channel selectively in this cell-type, and showed that direct recruitment of this otherwise genetically-inaccessible population elicits behavioral aversion. The specificity and minute-resolution of FLiCRE enables molecularly-informed characterization, manipulation, and reprogramming of activated cellular ensembles. Kim et al. present FLiCRE (Fast Light and Calcium-Regulated Expression), a stable, genetically engineered tool that enables activity-dependent tagging of cell populations within minutes in the mouse brain. Here, FLiCRE is used to identify and further characterize transiently activated neurons in the nucleus accumbens involved in driving behavioral aversion.
登录
查看更多内容
影响因子：
25
作者：
Hrvatin S;Hochbaum DR;Nagy MA;Cicconet M;Robertson K;Cheadle L;Zilionis R;Ratner A;Borges-Monroy R;Klein AM;Sabatini BL;Greenberg ME
通讯作者：
Greenberg ME
影响因子：
16.2
作者：
Burnett CJ;Li C;Webber E;Tsaousidou E;Xue SY;Brüning JC;Krashes MJ
通讯作者：
Krashes MJ
影响因子：
25
作者：
DeNardo, Laura A.;Liu, Cindy D.;Luo, Liqun
通讯作者：
Luo, Liqun
影响因子：
25
作者：
Boyden, ES;Zhang, F;Deisseroth, K
通讯作者：
Deisseroth, K
影响因子：
64.8
作者：
Jennings, Joshua H.;Kim, Christina K.;Deisseroth, Karl
通讯作者：
Deisseroth, Karl