Pseudorabies Virus (PRV) for DRG retrograde labeling?

viral_vectors

#1

Anyone have thoughts or experiences with this?


#2

I’m not sure it would adequately label DRG and afferents. In our experience, we injected PRV-152 (Bartha strain expressing PRV) into the hindlimb muscle. We saw robust infection of spinal cord motor neurons but not substantial DRG infection. Moreover we definitely did not see GFP-labeled afferents in the spinal cord.

Our study relied heavily on previous work from the following paper, which is a useful reference on PRV-152:

Also, it’s worth mentioning that the PRV vectors described in this paper are attenuated but replication competent. They will move transsynaptically in the retrograde direction and (in our experience) will eventually kill the animal around the time the infection reaches the brain. Also, the health of infected neurons is not great unless you catch them very soon after they start expressing the GFP.

A subsequent publication describes a PRV-cre vector (Becker strain) which is replication deficient, improving safety and animal health, and limiting gene expression to only the primarily infected neurons and not their presynaptic partners.

In our hands this vector did not result in detectable cre expression at either the site of injection or the spinal projection neurons we were attempting to label. However, this may not reflect the capabilities of this vector in a R26-CAG-LSL-reporter mouse where only a small amount of cre is necessary to enable strong expression of a reporter. We were using this vector in neonatal rat, in conjunction with a second cre-dependent AAV, so it is also possible that our lack of success was due to the cre-dependent vector and not the PRV-cre.


#3

Wow. Thanks @liz! This is extremely helpful. Thanks for sharing. I may pass on PRV. The quest continues for a good anterograde transsynaptic tracer that can start with DRG neurons…