Xiaobu tips | Peripheral PRV tracer - fast, accurate and bright! Everything you want is here!

The brain is the most complex system in the body, determining physiological activities such as emotion, memory, and learning. Neural circuits are the basis for brain functions. Clarifying the composition and working principles of neural circuits is of great significance for understanding, utilizing and protecting the brain. Currently, neurotropic viruses are widely used in anatomical research on central and peripheral nerves and play an important role in the analysis of neural circuit structure and function.

PRV Bartha was originally used to study efferent nerve pathways by injecting peripheral tissue, and to study peripheral nerve pathways through antibody immunohistochemistry. After intravitreal injection of PRV-152, a PRV Bartha derivative that expresses green fluorescent protein, in hamsters, the virus only spreads retrogradely across synapses through autonomic nervous circuits and infects the suprachiasmatic nucleus [1]. Banfield[2] and others constructed the PRV Bartha recombinant virus PRV-614 expressing red fluorescent protein, established a dual-virus trans-neuron labeling technology, and studied the connection between a single neuron and multiple neural circuits in the brain. Jia et al. [3] optimized the expression level of the reporter gene and constructed two new retrograde trans-polysynaptic tracer viruses, PRV531 and PRV724.

With the widespread application of PRV viruses in the research fields of central nervous circuits and central regulatory networks of peripheral organs, Brinkes has accumulated rich experience in PRV production and use. At present, the use of PRV in central nervous system circuit tracing research is becoming mature. However, in the analysis of the central regulatory network of peripheral organs, there are always problems with poor tracing effects, such as unstable tracing results and weak fluorescence signals. , low marking efficiency, etc. For details, please see previous tweets: Xiaobu tips | Peripheral-central nervous circuit marking overall service

Below, Xiaobu will share some tips for using PRV using the injection test results of our company's PRV at central markers and different peripheral parts as examples.

Comparison of central and peripheral use of PRV in mice

Picture of the tracer test effect of PRV in the central and peripheral mice

1. Central network tracer

Product: BC-PRV-724;
Dose: 100 nl;
Expression time: 3 days.

2. Tracing the neural regulatory network of a certain internal organ

Product: BC-PRV-531-Plus;
dosage: 3 ul;
expression time: 4-5 days.

3. Tracing the neural regulatory network of a certain gland

Product: BC-PRV-531-Pro;
Dosage: 3 ul;
Expression time: 4-5 days.

Notes:

• 1. Since the PRV virus has a certain toxic effect on the animal body, it is recommended to select adult and healthy animals to conduct experiments in order to complete relevant tracer research more efficiently.
• 2. After the PRV virus is injected, the mice will show corresponding symptoms in about 3-5 days. The status of the mice should be monitored regularly. When symptoms such as curling up, twitching, and trembling appear, perfusion can be performed. When the mice are in a dying state, samples can be collected. PRV infection works best.
• 3. After injection of PRV virus, the time window for mice to develop symptoms is difficult to control. It is recommended to increase the frequency of monitoring mice from the third day after injection;
• 4. PRV is a reverse trans-polysynaptic virus. During animal experiments, different sampling times can be set according to the number of loop connections. According to the life cycle of the virus, it is estimated that its propagation speed in the center may be 24-36 hours across the first-level synapses; 36-48 hours across the two-level synapses; 60-72 hours across the third-level synapses or more, so it should be based on experimental needs. Arrange sampling time reasonably.
• 5. In in vivo experiments, if the fluorescence signal is weak due to short expression time and other reasons, the fluorescence signal can be amplified through immunohistochemistry experiments.

PRV related products and services

references

• 1. Pickard GE, Smeraski CA, Tomlinson CC, Banfield BW, Kaufman J, Wilcox CL, Enquist
• LW, Sollars P JJ Neurosci., 2002, 22: 2701-2710
• 2. Banfield BW, Kaufman JD, Randall JA, Pickard GE. J. Virol. , 2003, 77: 10106-10112
• 3. Jia F, Lv P, Miao H, Shi X, Mei H, Li L, Xu X, Tao S, Xu F．Front. Neuroanat., 2019, 13: 63

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