An anterograde rabies virus vector for high-resolution large-scale reconstruction of 3D neuron morphology.

Matthias Georg Haberl,Silvia Viana Da Silva,Christophe Mulle,Andreas Frick,Marcel Oberlaender,Alexander Ghanem,Jason M Guest,Melanie Ginger,Karl-Klaus Conzelmann

doi:10.1007/s00429-014-0730-z

Abstract

Glycoprotein-deleted rabies virus (RABV ∆G) is a powerful tool for the analysis of neural circuits. Here, we demonstrate the utility of an anterograde RABV ∆G variant for novel neuroanatomical approaches involving either bulk or sparse neuronal populations. This technology exploits the unique features of RABV ∆G vectors, namely autonomous, rapid high-level expression of transgenes, and limited cytotoxicity. Our vector permits the unambiguous long-range and fine-scale tracing of the entire axonal arbor of individual neurons throughout the brain. Notably, this level of labeling can be achieved following infection with a single viral particle. The vector is effective over a range of ages (>14 months) aiding the studies of neurodegenerative disorders or aging, and infects numerous cell types in all brain regions tested. Lastly, it can also be readily combined with retrograde RABV ∆G variants. Together with other modern technologies, this tool provides new possibilities for the investigation of the anatomy and physiology of neural circuits.

Highlights

The reconstruction of neuronal circuits is central to many questions in neuroscience
To render rabies virus (RABV) dentate gyrus (DG) capable of cell body infection, we pseudotyped it with a chimeric envelope protein containing the N-terminal domain of the vesicular stomatitis virus glycoprotein (VSV-G)
VSV-G binds to highly ubiquitous receptors (Finkelshtein et al 2013), conferring the ability to transduce a wide range of cell types, a property that has previously been exploited for the production of VSV-G pseudotyped viral vectors such as retro- and lentiviruses (Burns et al 1993)

Summary

Introduction

The reconstruction of neuronal circuits is central to many questions in neuroscience. Knowledge of the finescale morphology of neurons provides insight into the identity and function of individual neurons, and into the function of neural circuits S. Viana da Silva PDBEB CNC, University of Coimbra, Coimbra, Portugal. Oberlaender Digital Neuroanatomy, Max Planck Florida Institute for Neuroscience, Jupiter, FL, USA

Methods

Results

Conclusion