Viral Transneuronal Tracing Technology: Defining the Synaptic Organisation of Neural Circuits

Abstract

Viral transneuronal tracing is a powerful approach that has been widely used to define the cellular organisation of functionally defined systems that underlie brain function. The method exploits the ability of neurotropic viruses to infect neurons and produce infectious progeny that pass through the intimate synaptic connections among neurons to infect polysynaptic pathways. A number of important technological advances in recent years have dramatically improved the ability of this experimental approach to define the synaptology of complex neural circuits. In this chapter we provide an overview of the method as well as mechanisms that contribute to its effectiveness for circuit analysis. We focus on alpha herpesviruses and rabies virus as these pathogens have been most widely applied in transneuronal tracing studies.

Key Concepts:

  • Viral transneuronal tracing exploits the invasive properties of neurotropic viruses.

  • Knowledge of the functional organisation of the viral genome is fundamental to an informed application of the viral transneuronal tracing method.

  • Circuit architecture is influential in determining the progression of viral infection through a neural circuit.

  • Isogenic recombinant viruses expressing unique reporters permit analysis of collateralisation of projections within complex neural circuits.

  • Colocalisation of reporters of infection with neurotransmitters and peptides permits neurochemical phenotyping of neurons within labelled circuits.

Keywords: alpha herpesvirus; pseudorabies; rabies; transneuronal; transsynaptic

Figure 1.

The attributes of ‘classical’ versus viral transneuronal tracing technology are illustrated. Both of these tracing approaches exploit axonal transport capabilities of neurons to establish connections between regions and among neurons. Similarly, both classes of tracers can be separated into different classes based on the direction of transport. In either case, tracers are taken up by either by the cell bodies or axon terminals. Transport from the cell body to the terminal is termed ‘anterograde’ transport whereas transport from axon terminals to the soma is termed ‘retrograde’ transport. The major distinction between classical tracers and viral tracers relates to the extent of labelling of a circuit. Classical tracers (with few exceptions) do not cross synapses and therefore only establish regional associations. In contrast, viral transneuronal tracers cross synapses and, with advancing time, sequentially infect synaptically linked neurons. The replication of virus in each neuron amplifies the signal produced within neurons of a circuit, aiding in their localisation in the brain.

Figure 2.

The life cycles of pseudorabies virus and rabies virus, and their utility for circuit analysis, are illustrated schematically. In spite of the similarity in their common names, the two viruses use distinctly different mechanisms for replication and transneuronal infection. The major distinction in this regard is the fact that pseudorabies virus is a DNA virus and rabies virus is an RNA virus. The major steps in the life cycle of each virus are illustrated and labelled. See the text for a more detailed description.

Figure 3.

The unique advantages of viral transneuronal labelling technology is illustrated in this schematic diagram of the polysynaptic organisation of circuitry revealed by transport of rabies virus through primate neural networks following injection of rabies virus into motor cortex (M1) of nonhuman primate brain. Progressive retrograde transneuronal transport of the virus through multiple synapses reveals all cell groups synaptically linked to M1, including cerebellar components of the network. Reproduced with permission from Kelly and Strick .

Figure 4.

Retrograde transneuronal infection of Purkinje neurons in nonhuman primate brain following injection of rabies virus into motor cortex are illustrated. See Figure for the organisation of the circuitry that leads to this labelling. Immunocytochemical localisation of viral antigens reveals neurons infected by transneuronal passage of virus. Note that vial antigen is present throughout the somatodendritic compartments of neurons, revealing the cytoarchitecture of the infected neurons and further demonstrating that virus passes transneuronally from even the most distal branches of the dendritic tree. Reproduced with permission from Kelly and Strick .

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Strick, Peter L, and Card, J Patrick(Dec 2011) Viral Transneuronal Tracing Technology: Defining the Synaptic Organisation of Neural Circuits. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1002/9780470015902.a0022359]