Correia Patrícia A, Matias Sara, Mainen Zachary F
Champalimaud Research, Champalimaud Centre for the Unknown, Lisbon, Portugal.
Bio Protoc. 2017 Sep 20;7(18):e2549. doi: 10.21769/BioProtoc.2549.
Optogenetic methods are now widespread in neuroscience research. Here we present a detailed surgical procedure to inject adeno-associated viruses and implant optic fiber cannulas in the dorsal raphe nucleus (DRN) of living mice. Combined with transgenic mouse lines, this protocol allows specific targeting of serotonin-producing neurons in the brain. It includes fixing a mouse in a stereotaxic frame, performing a craniotomy, virus injection and fiber implantation. Animals can be later used in behavioral experiments, combined with optogenetic manipulations (Dugué ., 2014; Correia ., 2017) or monitoring of neuronal activity (Matias ., 2017). The described procedure is a fundamental step in both optogenetic and fiber photometry experiments of deep brain areas. It is optimized for serotonin neurons in the DRN, but it can be applied to any other cell type and brain region. When using transgenic mouse lines that express functionally relevant levels of optogenetic tools or reporter lines, the virus injection step can be skipped and the protocol is reduced to the cannula implantation procedure.
光遗传学方法如今在神经科学研究中广泛应用。在此，我们展示一种详细的手术程序，用于在活体小鼠的中缝背核（DRN）中注射腺相关病毒并植入光纤套管。结合转基因小鼠品系，该方案能够特异性靶向大脑中产生血清素的神经元。它包括将小鼠固定在立体定位框架中、进行开颅手术、病毒注射和光纤植入。之后可将动物用于行为实验，结合光遗传学操作（Dugué 等人，2014年；Correia 等人，2017年）或监测神经元活动（Matias 等人，2017年）。所描述的程序是深部脑区光遗传学和光纤光度学实验的基本步骤。它针对DRN中的血清素神经元进行了优化，但也可应用于任何其他细胞类型和脑区。当使用表达功能相关水平的光遗传学工具或报告基因品系的转基因小鼠品系时，可跳过病毒注射步骤，该方案简化为套管植入程序。
