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

Haberl, Matthias G.; Silva, Silvia Viana da; Guest, Jason Mike; Ginger, Melanie; Ghanem, Alexander; Mulle, Christophe; Oberlaender, Marcel; Conzelmann, Karl‐Klaus; Frick, Andreas

doi:10.1007/s00429-014-0730-z

Cited by 31 publications

(27 citation statements)

References 47 publications

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“…Infection by the VSV G-coated viruses was found restricted to tight cluster of cells directly at the injection sites (Haberl et al, 2014;Wickersham et al, 2013), while RABV G-complemented viruses stained cells sending axonal inputs to the injection site (Fig. 3).…”

Section: Labeling Of Neurons With Pseudotyped G Rabv By Retro-or Antementioning

confidence: 96%

“…Infection with a single RABV virus is sufficient to initiate an infectious cycle and full fluorescent protein gene expression, the use of limiting MOI therefore allows sparse labeling and detailed morphological analysis of neurons both in vitro and in vivo (Haberl et al, 2014). Pseudotyping of a G gene-deficient RABV with the cognate RABV G protein obviously preserves the natural features of RABV in that the viruses enters neurons at axon terminals, and is transported with p75NTR-containing vesicles to the cell body where it can enter the cytoplasm and initiate an infectious cycle Klingen et al, 2008).…”

Section: Labeling Of Neurons With Pseudotyped G Rabv By Retro-or Antementioning

confidence: 99%

“…The recent identification of the almost ubiquitous low density lipoprotein receptor (LDLR) family members as receptors for VSV (Finkelshtein et al, 2013) may explain this extended tropism. In fact, G RABV complemented with a VSV Indiana G construct containing the RABV SAD B19 G C-tail or both transmembrane and C-tail (Haberl et al, 2014), have obviously lost the retrograde infection route and have gained an extended tropism, including astrocytes.…”

Section: Labeling Of Neurons With Pseudotyped G Rabv By Retro-or Antementioning

confidence: 99%

“…Since RABV (VSV G) accepts the insertion of large and multiple genes, the latter represents a promising anterograde viral vector for applications like single neuron reconstruction. The availability of retrograde (RABV G) and anterograde (VSV G) tracers with identical kinetics and performance also allows reciprocal tracing of projections to and from a region of interest (Haberl et al, 2014;Wickersham et al, 2013).…”

Section: Labeling Of Neurons With Pseudotyped G Rabv By Retro-or Antementioning

confidence: 99%

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G gene-deficient single-round rabies viruses for neuronal circuit analysis

Ghanem

Conzelmann

2016

Virus Research

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Section: Labeling Of Neurons With Pseudotyped G Rabv By Retro-or Antementioning

confidence: 96%

Section: Labeling Of Neurons With Pseudotyped G Rabv By Retro-or Antementioning

confidence: 99%

Section: Labeling Of Neurons With Pseudotyped G Rabv By Retro-or Antementioning

confidence: 99%

Section: Labeling Of Neurons With Pseudotyped G Rabv By Retro-or Antementioning

confidence: 99%

See 2 more Smart Citations

G gene-deficient single-round rabies viruses for neuronal circuit analysis

Ghanem

Conzelmann

2016

Virus Research

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“…Monosynaptically restricted transsynaptic tracing by green fluorescent protein (GFP) expressing G gene-deleted rabies viruses (20,21) represented a pioneering step in neurobiology, as the system allowed reliable tracing of retrograde transsynaptic neuron connections. Similarly to the case for RABV, which was also modified to allow nonretrograde infection by introduction of vesicular stomatitis virus (VSV) G sequences (22,23), the system was also adapted to VSV vectors for use as retro-and anterograde neuronal tracers in a G-dependent manner (16,24).…”

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confidence: 99%

Anterograde Glycoprotein-Dependent Transport of Newly Generated Rabies Virus in Dorsal Root Ganglion Neurons

Bauer

Nolden

Schröter

et al. 2014

J Virol

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Rabies virus (RABV) spread is widely accepted to occur only by retrograde axonal transport. However, examples of anterograde RABV spread in peripheral neurons such as dorsal root ganglion (DRG) neurons indicated a possible bidirectional transport by an uncharacterized mechanism. Here, we analyzed the axonal transport of fluorescence-labeled RABV in DRG neurons by livecell microscopy. Both entry-related retrograde transport of RABV after infection at axon endings and postreplicative transport of newly formed virus were visualized in compartmentalized DRG neuron cultures. Whereas entry-related transport at 1.5 m/s occurred only retrogradely, after 2 days of infection, multiple particles were observed in axons moving in both the anterograde and retrograde directions. The dynamics of postreplicative retrograde transport (1.6 m/s) were similar to those of entry-related retrograde transport. In contrast, anterograde particle transport at 3.4 m/s was faster, indicating active particle transport. Interestingly, RABV missing the glycoproteins did not move anterogradely within the axon. Thus, anterograde RABV particle transport depended on the RABV glycoprotein. Moreover, colocalization of green fluorescent protein (GFP)-labeled ribonucleoproteins (RNPs) and glycoprotein in distal axonal regions as well as cotransport of labeled RNPs with membrane-anchored mCherry reporter confirmed that either complete enveloped virus particles or vesicle associated RNPs were transported. Our data show that anterograde RABV movement in peripheral DRG neurons occurs by active motor protein-dependent transport. We propose two models for postreplicative long-distance transport in peripheral neurons: either transport of complete virus particles or cotransport of RNPs and G-containing vesicles through axons to release virus at distal sites of infected DRG neurons. IMPORTANCERabies virus retrograde axonal transport by dynein motors supports virus spread over long distances and lethal infection of the central nervous system. Though active rabies virus transport has been widely accepted to be unidirectional, evidence for anterograde spread in peripheral neurons supports the hypothesis that in some neurons RABV also enters the anterograde pathway by so-far unknown mechanisms. By live microscopy we visualized fast anterograde axonal transport of rabies virus. The velocities exceeded those of retrograde movements, suggesting that active, most likely kinesin-dependent transport machineries are involved. Dependency of anterograde transport on the expression of virus glycoprotein G and cotransport with vesicles further suggest that complete enveloped virus particles or cotransport of virus ribonucleoprotein and G-containing vesicles occurred. These data provide the first insight in the mechanism of anterograde rabies virus transport and substantially contribute to the understanding of RABV replication and spread of newly formed virus in peripheral neurons.

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Laminar specific gene expression reveals differences in postnatal laminar maturation in mouse auditory, visual, and somatosensory cortex

Chang¹,

Suzuki²,

Kawai³

2018

J of Comparative Neurology

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Proper formation of laminar structures in sensory cortexes is critical for sensory information processing. Previous studies suggested that the timing of neuronal migration and the laminar position of cortical neurons differ among sensory cortexes. How they differ during postnatal development has not been systematically investigated. Here, identifying laminas using transcription factors, we examined postnatal changes in neuronal density and distribution in presumptive primary auditory (ACx), visual (VCx), and somatosensory cortexes (SCx) in a strain of mice using immunofluorescence techniques. Development of laminar thickness and its cortical proportion differed among the sensory cortexes. Layers 2-4 defined by Cut-like homeobox 1 (Cux1)-expressing neurons were narrower, and layer 5 was wider in ACx compared to those in VCx or SCx, while Forkhead-box protein P2 (Foxp2)-defined layer 6 was wider in SCx than the other two sensory cortexes throughout postnatal development. Meanwhile, thalamocortical input layers identified by Cux1-expressing neurons formed later in ACx than in the other two cortical regions. The cell densities of ETS-related protein 81-expressing neurons increased in both lower and upper layers but at distinct timing, while those of COUP-TF-interacting protein 2 expressing neurons in the lower layers changed bidirectionally (i.e., increased or decreased) both in layer- and cortical region-specific manners. Foxp2-expressing cells in layer 6 distributed differently and declined at different timing among the sensory cortexes. Overall, we demonstrate that the maturational timing of lamina differs among the sensory cortexes and that postnatal age-dependent changes in neuronal distribution are unique to each of the sensory cortexes.

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

Cited by 31 publications

References 47 publications

G gene-deficient single-round rabies viruses for neuronal circuit analysis

G gene-deficient single-round rabies viruses for neuronal circuit analysis

Anterograde Glycoprotein-Dependent Transport of Newly Generated Rabies Virus in Dorsal Root Ganglion Neurons

Laminar specific gene expression reveals differences in postnatal laminar maturation in mouse auditory, visual, and somatosensory cortex

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