nIRCat characterization and acute brain slice imaging

Dopamine neuromodulation of neural synapses is a process implicated in a number of critical brain functions and diseases. Development of protocols to visualize this dynamic process is essential to understanding how dopamine modulates neurochemical brain function. We have developed a non-genetically encoded, near infrared (nIR) catecholamine nanosensor (nIRCat) capable of identifying ~2 µm dopamine release hotspots in the dorsal striatal brain slices. This platform is readily introduced into both genetically tractable and intractable organisms and is compatible with a number of dopamine receptor agonist and antagonists. In the following work, we describe the synthesis, characterization, and implementation of nIRCat in brain slices. We show how to image electrically and optogenetically stimulated dopamine release using nIRCat, and how these imaging protocols can be adapted to study the effects of dopamine receptor pharmacology. We also detail the development of video-analysis software to ...

多巴胺对神经突触的神经调节作用是一种与多种关键脑功能及疾病密切相关的过程。开发可视化这一动态过程的实验方案，对于理解多巴胺如何调控脑内神经化学功能至关重要。我们开发了一种非基因编码的近红外（near infrared, nIR）儿茶酚胺纳米传感器（nIRCat），能够在背侧纹状体脑切片中识别出约2微米的多巴胺释放热点。该平台可轻松应用于基因可操作及不可操作的生物体系，且与多种多巴胺受体激动剂和拮抗剂兼容。在后续研究中，我们详细描述了nIRCat的合成、表征及其在脑切片中的应用方法。我们展示了如何利用nIRCat对电刺激及光遗传学（optogenetic）刺激诱导的多巴胺释放进行成像，以及如何调整这些成像方案以研究多巴胺受体药理学效应。我们还详细介绍了视频分析软件的开发，以用于……