PRV Neural Circuit Tracing Services, Supporting Peripheral-Central Research
Release time:2024-09-26 16:18:37
1. What is Pseudorabies Virus (PRV)?
Pseudorabies virus (PRV), also known as porcine herpesvirus type 1, has a genome of approximately 150 kb linear double-stranded DNA. The Bartha strain of PRV exhibits retrogradetrans-synaptic properties. Recombinant tracing tool viruses based on the Bartha strain can strictly label neural networks through multi-level retrograde trans-synaptic pathways. After infecting neural cells, the virus replicates intracellularly and expresses target genes, with progeny viruses transporting across synapses to enter upstream neurons, initiating a new round of replication and retrograde trans-synaptic spread.
2. What can PRV do in neural circuit research?
PRV is significant for studies related to peripheral-central and central-central pathways, often used to explore central nervous nuclei directly or indirectly associated with specific peripheral organs. For example, to identify which brain nuclei initiate or participate in commands to lift the thigh, PRV can be injected into the target muscle group, followed by brain tissue sampling after a few days to observe fluorescent signals in central brain slices, identifying nuclei involved in thigh lifting.
Case 1:
·Objective: Investigate the neural circuits related to thigh lifting and assess the differences in neural regulation between left and right limbs.
·Protocol: Use PRV tool viruses to inject 2.5 μl of PRV-803 (red) into the left thigh muscle and PRV-531 (green) into the right thigh muscle of mice, ensuring skin is cut and sutured post-injection. Observe the animals at least twice daily for 1-3 days to ensure normal recovery post-surgery; from the third day after injection, increase monitoring frequency to no longer than every 4 hours.
Figure 1: Schematic Diagram of Virus Injection
·Results: Approximately five days post-injection, the animals exhibited noticeable neurological symptoms, prompting us to perform cardiac perfusion and brain sampling. After freezing and slicing the brain tissue, we obtained the following data. Based on the results, we believe there are direct or indirect connections between the control of the target muscle group and nuclei such as the motor cortex (MOp), the suprachiasmatic nucleus (SCN), and the periaqueductal gray (PAG). Further validation of the significance of these nuclei can be conducted using calcium signaling recordings or by employing optogenetic or chemogenetic techniques to manipulate these nuclei, ultimately identifying central nuclei more closely related to the target muscle group.
Figure 2: Retrograde Multi-Level Virus Tracing of Muscles (Brain Case Test Data)
Case 2:
·Experimental Objective: To investigate the neural circuits associated with and controlling urination, specifically which key brain regions are involved in the body's autonomous urination.
·Experimental Plan: The PRV (pseudorabies virus) tracer virus was injected in small amounts into the left and right sides of the bladder wall using PRV-531 (green) and PRV-724 (red); due to significant neurological symptoms and eventual death in animals infected with PRV, the animals were closely monitored daily to ensure recovery to a normal state post-surgery. When in a near-death condition, the animals were subjected to cardiac perfusion to extract the brain.
·Experimental Results: About five days after the virus injection, the animals exhibited notable neurological symptoms. Researchers gradually performed cardiac perfusion to extract the brain based on the observed conditions. Following the preparation of frozen sections of brain tissue, the following data was obtained. Based on the experimental results, researchers found that the primary motor cortex (M1), primary somatosensory cortex (S1), and the micturition center (PMC) have direct or indirect connections.
Figure 3: Retrograde multilevel viral tracing of bladder wall nerves (PMID: 30361547)
3. PRV Neural Circuit Tracing Services Offered by Brain Case
In recent years, the research focus on peripheral-central connections has surged, with more peripheral organs becoming the subject of investigation. Brain Case is committed to customer-oriented services, supporting research on peripheral-central neural circuits. We provide a comprehensive PRV neural circuit tracing service, including the stable older versions PRV531 (green) and PRV724 (red), as well as the efficient and bright new products PRV801 (green) and PRV803 (red), which have been successfully applied in multiple high-interest peripheral tissue studies. Below are examples of research sites we have operated on:
◆ Digestive System: Duodenum, cecum, colon, rectum, pancreas
◆ Endocrine System: Thyroid
◆ Reproductive System: Testes, ovaries, epididymis, prostate
◆ Urinary System: Bladder wall
◆ Musculoskeletal System: Muscle, bone marrow cavity
◆ Cardiovascular System: Heart
◆ Immune System: Spleen
◆ Circulatory System: Femoral cavity
◆ Nervous System: Brain + spinal cord
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