Table_2_Monosynaptic Tracing Success Depends Critically on Helper Virus Concentrations.XLSX

Monosynaptically-restricted transsynaptic tracing using deletion-mutant rabies virus (RV) has become a widely used technique in neuroscience, allowing identification, imaging, and manipulation of neurons directly presynaptic to a starting neuronal population. Its most common implementation is to use Cre mouse lines in combination with Cre-dependent “helper” adeno-associated viral vectors (AAVs) to supply the required genes to the targeted population before subsequent injection of a first-generation (ΔG) rabies viral vector. Here we show that the efficiency of transsynaptic spread and the degree of nonspecific labeling in wild-type control animals depend strongly on the concentrations of these helper AAVs. Our results suggest practical guidelines for achieving good results.

使用缺失突变型狂犬病病毒（rabies virus, RV）的单突触限制性跨突触追踪技术，现已成为神经科学领域应用广泛的研究手段，可对起始神经元群体的直接突触前神经元进行鉴定、成像与操控。该技术最常用的实施策略为：将Cre小鼠品系与依赖Cre的“辅助”腺相关病毒载体（adeno-associated viral vectors, AAVs）联用，在后续注射第一代（ΔG）狂犬病病毒载体之前，向靶神经元群体递送所需基因。本研究证实，野生型对照动物中的跨突触传播效率与非特异性标记程度，显著依赖于上述辅助AAV的浓度。本研究结果可为获得理想实验结果提供实用操作指南。