Retrograde Muti-synaptic Tracing-PRV-Brain Case ()

Peripheral to Central, a tool for dissecting the motor output circuits - PRV

The wild-type strain of PRV (Becker strain) exhibits higher toxicity and can propagate both anterogradely and retrogradely during neural circuit transmission. In contrast, the vaccine strain of PRV (Bartha strain) has lower toxicity. Recombinant tracing tool viruses based on the PRV Bartha strain can strictly label neural networks in a retrograde multisynaptic manner. After infecting neural cells, the virus replicates and expresses the target genes intracellularly. The progeny viruses are transported to the synapse, where they undergo retrograde transsynaptic transmission to upstream neurons, initiating a new cycle of replication and retrograde transsynaptic propagation.

Figure 1. PRVBartha strain reverse transpolysynaptic labeling

48 hours after injection of PRV-hUbC-EGFP into the olfactory bulb (TU), retrograde multisynaptic labeling was observed in upstream brain regions, including M1, M2, Aco, Piri, OB, VTA, indicating direct or indirect projections from these areas to TU.

Figure 2. Injecting PRV into brain region TU can label input neural circuits of different orders in the injection area

application

1. Studying the multi-synaptic input network structure across the entire brain of specific regions.; (see case presentation 1)
2. Studying the multi-synaptic input network of two specific brain regions in the whole brain; (see case presentation 3)
3. Neural network changes during neural development;
4. In neuropsychiatric disease models, the structural characteristics and abnormal changes of the brain’s central and peripheral information output neural networks;
5. In the neural injury and rehabilitation model, the degree of damage and recovery of the connection status of the brain center and peripheral information output neural networks;
6. Tracing the motor output circuits between peripheral organs and the central nervous system. (See case presentation 2)

Case display

Example 1: Retrograde Muti-synaptic Tracing of DS

Experimental animals : C57BL/6Jmice
Virus used : PRV152-GFP
Experimental methods and results : PRV152-GFP injected into the bilateral dorsal striatum (DS) of mice, revealed retrograde transsynaptic spread to R-NG, DMV, NTS, AP, PBNdl, and PBNm. These findings further validate the existence of the R-NG→PBNdl→SNc pathway, consistent with previous results.

Figure 3. Validation of R-NG→PBNdl→SNc pathway (HanW, et al., Cell, 2018)

Example 2: Retrograde Muti-synaptic Tracing of bladder wall nerves

Experimental animals : C57BL/6Jmice
Viruses used : PRV-EGFP, PRV-RFP
Experimental methods and results : After injecting PRV-EGFP into the bladder wall of mice, EGFP signals were observed in the cortical S1/M1L5 pyramidal neurons. Subsequently, PRV-EGFP and PRV-RFP were injected into the right and left bladder walls of mice, respectively, resulting in the co-expression of EGFP and RFP in the cortical S1/M1L5 pyramidal neurons. These findings indicate that PRV-EGFP and PRV-RFP retrogradely transsynaptically spread indiscriminately to the cortical S1/M1L5 pyramidal neurons, which serve as the upstream brain neurons projecting to the bladder wall.

Figure 4. Identification of the M1L5 pyramidal neuron population in the upper bladder cortex (YaoJ, et al., NatNeurosc, 2018)

Example 3: Studying the multi-level input network of two specific brain regions across the entire brain.

Experimental animals : C57BL/6 mice
Viruses used : RV152(GFP), PRV614(RFP)
Experimental methods and results : In C57BL/6 mice, PRV152 (GFP) and PRV614 (RFP) were injected into the olfactory bulbs on both sides, and the infection timeline was controlled simultaneously to detect the expression distribution of fluorescent proteins in the whole brain. Several neurons co-labeled with both colors of virus were observed in the locus coeruleus area, indicating that neurons in these brain regions are involved in regulating the information processing of both olfactory bulbs. Similarly, through this method, it is possible to elucidate the common upstream regulation of one or even multiple levels of two specific nuclei in the brain, that is, which types of neurons in those brain nuclei simultaneously control the functions of the other two brain regions.

Figure 5. Through PRV labeling and tracing, it was found that neurons in this nucleus simultaneously participate in regulating the information processing of both olfactory bulbs.

Product List

-Enhanced versions of PRV products are continuously being updated. Welcome to consult more products.

Product No.	Product Name	Fluorescent Protein Carried	Function
BC-PRV-531-Plus	PRV-CAG-EGFP	EGFP	Green, retrograde muti-synaptic tracing
BC-PRV-803-Plus	PRV-CAG-3Ms	mScarlet3	Red, retrograde muti-synaptic tracing

Neural Circuit Tracing Services

Labeling Type	Synaptic Specificity	Common Tools
Retrograde Tracing	Non-synaptic	AAV2/Re, AAV2/11, RV-ΔG-N2cG, CTB
Retrograde Monosynaptic Tracing	Monosynaptic	RV-EnvA-ΔG-XFP
Retrograde Mutisynaptic Tracing	Mutisynaptic	PRV-XFP, PRV-△TK-DIO-XFP (Cre-dependent)
Anterograde Tracing	Non-synaptic	AAV2/2, AAV2/8, AAV2/9
Anterograde Monosynaptic Tracing	Monosynaptic	AAV2/1, AAV2/9-mWGA, Hs06, H361
Anterograde Mutisynaptic Tracing	Mutisynaptic	HSV, VSV
Deliverables (for all services above): Full project report and raw imaging data