Monosynaptic Circuit Tracing with Glycoprotein-Deleted Rabies Viruses

Callaway, Edward M.; Luo, Liqun

doi:10.1523/jneurosci.0409-15.2015

Cited by 379 publications

(376 citation statements)

References 60 publications

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“…To investigate the regulation of CnF and PPN GlutNs we traced the sources of neuronal inputs to each structure, using the rabies based mono-synaptically restricted retrograde trans-synaptic circuit tracing ( 23,24 see Methods; Fig. 6).…”

Section: Resultsmentioning

confidence: 99%

“…We used a glycoprotein (G)-deleted rabies virus (RV dG ) 23 pseudotyped with the envelope glycoprotein EnvA to allow selective infection of glutamatergic cells via the TVA receptor. The TVA receptor was delivered together with the Rabies glycoprotein conditionally to glutamatergic cells, by injecting 200 – 300 nl AAVdj-EF1a-FLEX-GTB virus (helper virus, Salk Institute, visualized in green in Fig.…”

Section: Methodsmentioning

confidence: 99%

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Midbrain circuits that set locomotor speed and gait selection

Caggiano¹,

Leiras²,

Goñi-Erro³

et al. 2018

Nature

364

511

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Summary Locomotion is a fundamental motor function common to the animal kingdom. It is executed episodically and adapted to behavioural needs including exploration, requiring slow locomotion, and escaping behaviour, necessitating faster speeds. The control of these functions originates in brainstem structures although the neuronal substrate(s) supporting them are debated. Here, we show in mice that speed/gait selection are controlled by glutamatergic excitatory neurons (GlutNs) segregated in two distinct midbrain nuclei: the Cuneiform Nucleus (CnF) and the Pedunculopontine Nucleus (PPN). GlutNs in each of those two regions are sufficient for controlling slower alternating locomotor behavior but only GlutNs in the CnF are necessary for high-speed synchronous locomotion. Additionally, PPN- and CnF-GlutNs activation dynamics and their input and output connectivity matrices support explorative and escape locomotion, respectively. Our results identify dual regions in the midbrain that act in common to select context dependent locomotor behaviours.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Midbrain circuits that set locomotor speed and gait selection

Caggiano¹,

Leiras²,

Goñi-Erro³

et al. 2018

Nature

364

511

View full text Add to dashboard Cite

show abstract

“…Limitations of this technique are known (Callaway and Luo, 2015). Additionally, since only one in every four sections throughout the brain was mapped and analyzed, each distribution of input cells should be viewed as a sample of a sample for this viral tracing, as previously suggested (Ginger et al, 2013).…”

Section: Discussionmentioning

confidence: 99%

The Input-Output Relationship of the Cholinergic Basal Forebrain

2017

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Summary Basal forebrain cholinergic neurons influence cortical state, plasticity, learning and attention. They collectively innervate the entire cerebral cortex, differentially controlling acetylcholine efflux across different cortical areas and timescales. Such control might be achieved by differential inputs driving separable cholinergic outputs, although no input-output relationship on a brain-wide level has ever been demonstrated. Here we identify input neurons to cholinergic cells projecting to specific cortical regions by infecting cholinergic axon terminals with a monosynaptically-restricted viral tracer. This approach revealed several circuit motifs, such as central amygdala neurons synapsing onto basolateral amygdala-projecting cholinergic neurons or strong somatosensory cortical input to motor cortex-projecting cholinergic neurons. The presence of input cells in the parasympathetic midbrain nuclei contacting frontally projecting cholinergic neurons suggest that the network regulating the inner eye muscles are additionally regulating cortical state via acetylcholine efflux. This dataset enables future circuit-level experiments to identify drivers of known cholinergic cortical functions.

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“…The G-deleted rabies virus has been effective in driving EGFP expression in hundreds of retrogradely labeled neurons following injection into a target structure. The same rabies virus strain is similarly effective in multiple species including macaque monkeys 5,9 , ferrets (Hasse & Briggs, in revision), and rodents 7,8 . Accordingly, modified rabies virus can be utilized to selectively label the complete dendritic arborization patterns of any population of neurons following injection into a target structure.…”

Section: Discussionmentioning

confidence: 99%

Large-scale Reconstructions and Independent, Unbiased Clustering Based on Morphological Metrics to Classify Neurons in Selective Populations

Bragg

Briggs

2017

JoVE

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This protocol outlines large-scale reconstructions of neurons combined with the use of independent and unbiased clustering analyses to create a comprehensive survey of the morphological characteristics observed among a selective neuronal population. Combination of these techniques constitutes a novel approach for the collection and analysis of neuroanatomical data. Together, these techniques enable large-scale, and therefore more comprehensive, sampling of selective neuronal populations and establish unbiased quantitative methods for describing morphologically unique neuronal classes within a population.The protocol outlines the use of modified rabies virus to selectively label neurons. G-deleted rabies virus acts like a retrograde tracer following stereotaxic injection into a target brain structure of interest and serves as a vehicle for the delivery and expression of EGFP in neurons. Large numbers of neurons are infected using this technique and express GFP throughout their dendrites, producing "Golgi-like" complete fills of individual neurons. Accordingly, the virus-mediated retrograde tracing method improves upon traditional dye-based retrograde tracing techniques by producing complete intracellular fills.Individual well-isolated neurons spanning all regions of the brain area under study are selected for reconstruction in order to obtain a representative sample of neurons. The protocol outlines procedures to reconstruct cell bodies and complete dendritic arborization patterns of labeled neurons spanning multiple tissue sections. Morphological data, including positions of each neuron within the brain structure, are extracted for further analysis. Standard programming functions were utilized to perform independent cluster analyses and cluster evaluations based on morphological metrics. To verify the utility of these analyses, statistical evaluation of a cluster analysis performed on 160 neurons reconstructed in the thalamic reticular nucleus of the thalamus (TRN) of the macaque monkey was made. Both the original cluster analysis and the statistical evaluations performed here indicate that TRN neurons are separated into three subpopulations, each with unique morphological characteristics.

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Monosynaptic Circuit Tracing with Glycoprotein-Deleted Rabies Viruses

Abstract: Editor's Note: Toolboxes are intended to describe and evaluate methods that are becoming widely relevant to the neuroscience community or to provide a critical analysis of established techniques. For more information, see http://www.jneurosci.org/misc/ ifa_minireviews.dtl.

Cited by 379 publications

References 60 publications

Midbrain circuits that set locomotor speed and gait selection

Midbrain circuits that set locomotor speed and gait selection

The Input-Output Relationship of the Cholinergic Basal Forebrain

Large-scale Reconstructions and Independent, Unbiased Clustering Based on Morphological Metrics to Classify Neurons in Selective Populations

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