Department of Genetics and Department of Ophthalmology, Harvard Medical School, Harvard University and Howard Hughes Medical Institute Boston, MA, USA.
Front Neural Circuits. 2013 Feb 7;7:11. doi: 10.3389/fncir.2013.00011. eCollection 2013.
Defining the connections among neurons is critical to our understanding of the structure and function of the nervous system. Recombinant viruses engineered to transmit across synapses provide a powerful approach for the dissection of neuronal circuitry in vivo. We recently demonstrated that recombinant vesicular stomatitis virus (VSV) can be endowed with anterograde or retrograde transsynaptic tracing ability by providing the virus with different glycoproteins. Here we extend the characterization of the transmission and gene expression of recombinant VSV (rVSV) with the rabies virus glycoprotein (RABV-G), and provide examples of its activity relative to the anterograde transsynaptic tracer form of rVSV. rVSV with RABV-G was found to drive strong expression of transgenes and to spread rapidly from neuron to neuron in only a retrograde manner. Depending upon how the RABV-G was delivered, VSV served as a polysynaptic or monosynaptic tracer, or was able to define projections through axonal uptake and retrograde transport. In animals co-infected with rVSV in its anterograde form, rVSV with RABV-G could be used to begin to characterize the similarities and differences in connections to different areas. rVSV with RABV-G provides a flexible, rapid, and versatile tracing tool that complements the previously described VSV-based anterograde transsynaptic tracer.
定义神经元之间的连接对于理解神经系统的结构和功能至关重要。经过基因工程改造后能够在突触间传递的重组病毒为在体内解析神经元回路提供了一种强大的方法。我们最近证明，通过为病毒提供不同的糖蛋白，重组单纯疱疹病毒（VSV）可以获得顺行或逆行跨突触示踪能力。在这里，我们扩展了具有狂犬病毒糖蛋白（RABV-G）的重组 VSV（rVSV）的传输和基因表达的特征，并提供了其相对于顺行跨突触示踪 rVSV 的活性的示例。发现具有 RABV-G 的 rVSV 能够强烈表达转基因，并以逆行方式仅快速从神经元传播到神经元。根据 RABV-G 的递送方式，VSV 可作为多突触或单突触示踪剂，或者能够通过轴突摄取和逆行运输来定义投射。在同时感染顺行形式的 rVSV 的动物中，rVSV 与 RABV-G 可用于开始描述与不同区域的连接的相似和不同之处。具有 RABV-G 的 rVSV 提供了一种灵活、快速和通用的示踪工具，补充了先前描述的基于 VSV 的顺行跨突触示踪剂。
