Development of Pseudorabies Virus Strains Expressing Red Fluorescent Proteins: New Tools for Multisynaptic Labeling Applications

Banfield, Bruce W.; Kaufman, Jessica; Randall, Jessica A.; Pickard, Gary E.

doi:10.1128/jvi.77.18.10106-10112.2003

Cited by 120 publications

(131 citation statements)

References 22 publications

Supporting

Mentioning

125

Contrasting

Order By: Relevance

“…Genetically modified viral vectors like the PRV used in our study have greatly expanded our understanding of the intricate neural circuitry in the normal CNS (Banfield et al, 2003;Buijs et al, 2003;Card et al, 1990;Nadelhaft et al, 1992;Smith et al, 2000). Despite the important insights gained through the use of PRV, there are potential limitations to the utility of PRV as a transsynaptic marker, particularly in the complete SCI paradigm, in which the uptake of PRV by kidney-related sympathetic preganglionic neurons is markedly attenuated .…”

Section: Discussionmentioning

confidence: 99%

“…In addition, highly reactive OX-42-positive microglial cells and their processes were often observed in close association with preganglionic neurons in the thoracolumbar spinal cord at 96 h after inoculation of the kidney with PRV-614 (Fig. 6), a PRV Bartha construct that reports via a red fluorescent protein (Banfield et al, 2003).…”

Section: Microglial Phenotypes In the Spinal Cord After Injury And Nomentioning

confidence: 99%

“…The injections were uniformly placed at sites on the longitudinal midline of the convex surface of the kidney Tang et al, 2004). In some cases, PRV-614 (Banfield et al, 2003), which expresses monomeric red fluorescent protein (mRFP1; a generous gift of Roger Tsien) was used similarly. The injection sites were located by dividing the midline into thirds, and injecting at the rostral and caudal end of the middle third.…”

Section: Prv Injections Into the Kidneys Of Injured Rats Following Comentioning

confidence: 99%

See 2 more Smart Citations

Noxious Colorectal Distention in Spinalized Rats Reduces Pseudorabies Virus Labeling of Sympathetic Neurons

Duale

Lyttle²,

Smith³

et al. 2010

Journal of Neurotrauma

View full text Add to dashboard Cite

The retrograde transsynaptic tracer pseudorabies virus (PRV) has been widely used as a marker for synaptic connectivity in the spinal cord. Notably, the PRV-152 construct expresses enhanced green fluorescent protein (EGFP). We recently reported a significant attenuation of PRV-152 labeling of the intermediolateral cell column (IML) and celiac ganglia after complete T4 spinal cord transection versus sham injury in rats at 96 h after PRV-152 inoculation of the left kidney. Here we found a significant increase in noxious colorectal distention (CRD)-evoked c-Fos expression in spinal cords of injured versus sham rats without PRV infection. In order to assess whether enhancing neuronal activity in spinalized rats might increase PRV-152 labeling, we subjected awake spinalized rats to 1.5 h of intermittent noxious CRD either: (1) just prior to inoculation, or (2) 96 h after inoculation (n ¼ 3/group). Equal numbers of spinalized rats in both groups received PRV-152 inoculations without CRD (non-stimulated; n ¼ 3/group). At 96 h post-inoculation fixed spinal cords and left celiac ganglionic tissues were assessed for the distribution and quantification of EGFP-labeled cells. The injured cohort that received CRD just prior to PRV injection showed a significant reduction in EGFP-labeled cells in both the IML and left celiac ganglion compared to non-stimulated injured rats. In contrast, the injured cohort that received CRD 96 h after PRV-152 inoculation showed no differences in EGFP-labeled cell numbers in the IML or celiac ganglia versus non-stimulated injured rats. Interestingly, microglia near c-Fos-positive cells after acute CRD appeared more reactive compared to non-stimulated spinalized rats, and activated microglial cells markedly reduce viral transduction and progression following PRV inoculation of the CNS. Hence our results imply that increased CRD-induced c-Fos expression in the injured paradigm, prior to but not after PRV injection, further attenuates PRV-152 uptake, perhaps through changes in neuronal activity and/or innate neuro-immune responses.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Microglial Phenotypes In the Spinal Cord After Injury And Nomentioning

confidence: 99%

Section: Prv Injections Into the Kidneys Of Injured Rats Following Comentioning

confidence: 99%

See 1 more Smart Citation

Noxious Colorectal Distention in Spinalized Rats Reduces Pseudorabies Virus Labeling of Sympathetic Neurons

Duale

Lyttle²,

Smith³

et al. 2010

Journal of Neurotrauma

View full text Add to dashboard Cite

show abstract

“…In this study, we used two isogenic strains of the Bartha strain of PRV (PRV152 or PRV614) (Banfield et al, 2003), which were injected into the subcutaneous and abdominal fat depots of male and female rats. Different (red and green) fluorescent reporters associated with these viruses enables the identification of neurons at each synaptic relay that project through polysynaptic connections to either or both fat depots.…”

Section: Introductionmentioning

confidence: 99%

Neurochemical Characterization and Sexual Dimorphism of Projections from the Brain to Abdominal and Subcutaneous White Adipose Tissue in the Rat

et al. 2012

View full text Add to dashboard Cite

Retroperitoneal white adipose tissue (rWAT) and subcutaneous (inguinal) white adipose tissue (iWAT) are both innervated and regulated by sympathetic efferents, but the distribution and identity of the cells in the brain that regulate sympathetic outflow are poorly characterized. Our aim was to use two isogenic strains of a neurotropic virus (pseudorabies, Bartha) tagged with either green or red fluorescent reporters to identify cells in the brain that project to rWAT and/or iWAT. These viruses were injected into separate WAT depots in male and female Sprague Dawley rats. Retrogradely labeled neurons in the CNS were characterized by immunohistochemistry and PCR. For the latter, laser capture of individual virally labeled neurons was used. All virally labeled brain regions contained neurons projecting to either and both WAT depots. Neurons to abdominal fat were the most abundant in males, whereas females contained a greater proportion of neurons to subcutaneous via private lines and collateral branches. Retrogradely labeled neurons directed to WAT expressed estrogen receptor-␣ (ER␣), and fewer neurons to subcutaneous WAT expressed ER␣ in males. Regardless of sex, projections from the arcuate nucleus were predominantly from pro-opiomelanocortin cells, with a notable lack of projections from agouti-related protein-expressing neurons. Within the lateral hypothalamus, neurons directed to rWAT and iWAT expressed orexin and melaninconcentrating hormone (MCH), but male rats had a predominance of MCH directed to iWAT. In conclusion, the neurochemical substrates that project through polysynaptic pathways to iWAT and rWAT are different in male and female rats, suggesting that metabolic regulation of rWAT and iWAT is sexually dimorphic.

show abstract

“…PRV is neuron specific and passes transsynaptically to infect new neurons and renew the replication cycle (25). The Bartha strain used in this work is retrogradespecific, and expresses the GFP or mRed1 for visualization (PRV-152 or PRV-614, respectively) (26)(27)(28)(29)(30).…”

mentioning

confidence: 99%

Viral tracing identifies distributed columnar organization in the olfactory bulb

Willhite¹,

Nguyen²,

Masurkar³

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

157

142

View full text Add to dashboard Cite

Olfactory sensory neurons converge onto glomeruli in the olfactory bulb (OB) to form modular information processing units. Similar input modules are organized in translaminar columns for other sensory modalities. It has been less clear in the OB whether the initial modular organization relates to a columnar structure in the deeper layers involved in local circuit processing. To probe synaptic connectivity in the OB, we injected a retrograde-specific strain of the pseudorabies virus into the rat OB and piriform cortex. The viral-staining patterns revealed a striking columnar organization that extended across all layers of the OB from the glomeruli to the deep granule cell layer. We hypothesize that the columns represent an extension of the glomerular unit. Specific patterning was observed, suggesting selective, rather than distance-dependent, center-surround connectivity. The results provide a previously undescribed basis for interpreting the synaptic connections between mitral and granule cells within the context of a columnar organization in the OB and have implications for olfactory coding and network organization.columns ͉ olfaction ͉ pseudorabies virus ͉ synaptic tracing T he mammalian brain receives a diverse array of information about the external environment through highly varied sensory modalities. In sensory systems as dissimilar as somatosensation, vision, and audition, a common organizing principle has emerged within the sensory cortices, the cortical column. Lorente de Nó (1) first suggested a higher-order columnar organization of neurons based on Golgi impregnation anatomy of cortical cells. Electrophysiological evidence in the somatosensory cortex later showed that neurons responded to deep or superficial mechanical stimulation in grouped, vertical columns (2). This organizational principle was subsequently found in other sensory areas such as visual (3, 4) and auditory cortices (5) and in higher-order areas such as association cortex (6). The olfactory bulb (OB) is a cortical region for which there have been indications of possible columnar connectivity, but this organizational principle has not been explored as thoroughly in the OB as in other brain regions.In the olfactory system, the segregation of sensory information begins with the convergence of olfactory sensory neuron (OSN) axonal subpopulations onto specific glomeruli (Fig. 6, which is published as supporting information on the PNAS web site) (7). In rodents, the axons of 10,000 OSNs expressing the same olfactory receptor type in the nasal epithelium converge onto at least two glomeruli in each OB, one lateral and one medial, arising from OSNs in the medial and lateral epithelium, respectively (8-13). The glomerulus and the intrabulbar neurons synaptically connected within it are thought to represent a modular information processing unit that sends odor information to pyramidal neurons in the olfactory cortex and other brain areas via the projection neurons, mitral and tufted cells.Olfactory projection neurons receive inhibition at the gl...

show abstract

Development of Pseudorabies Virus Strains Expressing Red Fluorescent Proteins: New Tools for Multisynaptic Labeling Applications

Cited by 120 publications

References 22 publications

Noxious Colorectal Distention in Spinalized Rats Reduces Pseudorabies Virus Labeling of Sympathetic Neurons

Noxious Colorectal Distention in Spinalized Rats Reduces Pseudorabies Virus Labeling of Sympathetic Neurons

Neurochemical Characterization and Sexual Dimorphism of Projections from the Brain to Abdominal and Subcutaneous White Adipose Tissue in the Rat

Viral tracing identifies distributed columnar organization in the olfactory bulb

Contact Info

Product

Resources

About