Molecular profiling of neurons based on connectivity.
Molecular profiling of neurons based on connectivity.
复制标题
基于连通性的神经元分子分析。
DOI：
10.1016/j.cell.2014.03.059
复制
发表时间：
2014-05-22
期刊：
影响因子：
64.5
通讯作者：
Friedman JM
中科院分区：
文献类型：
--
作者：
Ekstrand MI;Nectow AR;Knight ZA;Latcha KN;Pomeranz LE;Friedman JM
神经回路的复杂性和细胞异质性对理解离散神经群体在控制行为中的作用提出了重大挑战。虽然神经解剖学方法能够实现神经回路的高分辨率绘图，但这些方法不允许基于神经元的连接性对神经元进行系统的分子分析。我们报告了一种对投射神经元进行分子分析的方法的发展。我们证明核糖体可以用针对 GFP 的骆驼纳米抗体进行标记，并且该系统可以被设计为选择性捕获来自用 GFP 逆行标记的神经元的翻译 mRNA。使用该系统，我们对投射到伏隔核的神经元进行了分析。然后，我们使用 AAV 选择性地分析投射到伏隔核的中脑多巴胺神经元。通过比较每个实验中捕获的 mRNA，我们鉴定了许多 VTA 多巴胺能投射神经元特异的标记。当前的方法提供了一种根据神经元的投影来分析神经元的方法。
The complexity and cellular heterogeneity of neural circuitry presents a major challenge to understanding the role of discrete neural populations in controlling behavior. While neuroanatomical methods enable high-resolution mapping of neural circuitry, these approaches do not allow systematic molecular profiling of neurons based on their connectivity. We report the development of an approach for molecularly profiling projective neurons. We show that ribosomes can be tagged with a camelid nanobody raised against GFP and that this system can be engineered to selectively capture translating mRNAs from neurons retrogradely labeled with GFP. Using this system we profiled neurons projecting to the nucleus accumbens. We then used an AAV to selectively profile midbrain dopamine neurons projecting to the nucleus accumbens. By comparing the captured mRNAs from each experiment, we identified a number of markers specific to VTA dopaminergic projection neurons. The current method provides a means for profiling neurons based on their projections.