Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, USA.
Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, USA.
Neuron. 2018 Jun 6;98(5):905-917.e5. doi: 10.1016/j.neuron.2018.05.028.
Retrogradely transported neurotropic viruses enable genetic access to neurons based on their long-range projections and have become indispensable tools for linking neural connectivity with function. A major limitation of viral techniques is that they rely on cell-type-specific molecules for uptake and transport. Consequently, viruses fail to infect variable subsets of neurons depending on the complement of surface receptors expressed (viral tropism). We report a receptor complementation strategy to overcome this by potentiating neurons for the infection of the virus of interest-in this case, canine adenovirus type-2 (CAV-2). We designed AAV vectors for expressing the coxsackievirus and adenovirus receptor (CAR) throughout candidate projection neurons. CAR expression greatly increased retrograde-labeling rates, which we demonstrate for several long-range projections, including some resistant to other retrograde-labeling techniques. Our results demonstrate a receptor complementation strategy to abrogate endogenous viral tropism and thereby facilitate efficient retrograde targeting for functional analysis of neural circuits.
逆行转运的神经嗜性病毒可根据其长程投射,实现对神经元的基因访问,已成为将神经连接与功能联系起来的不可或缺的工具。病毒技术的一个主要限制是,它们依赖于细胞类型特异性分子进行摄取和运输。因此,根据所表达的表面受体的互补性(病毒嗜性),病毒无法感染可变的神经元亚群。我们报告了一种受体互补策略,通过增强感兴趣病毒的感染能力来克服这一问题,在这种情况下,是犬腺病毒 2 型(CAV-2)。我们设计了 AAV 载体,在候选投射神经元中表达柯萨奇病毒和腺病毒受体(CAR)。CAR 表达大大提高了逆行标记率,我们在几种长程投射中证明了这一点,包括对其他逆行标记技术有抗性的投射。我们的结果证明了一种受体互补策略,可以消除内源性病毒嗜性,从而促进有效的逆行靶向,用于神经回路的功能分析。