Does any one have any thoughts about the performance of different AAV serotypes (and promoters) in the spinal dorsal horn?
My aim is to transduce dorsal horn neurons with intraspinal injections using a pan-neuronal promoter (Synapsin). I used AAV9 with a synapsin-driven gene and I saw really high transduction of deep dorsal horn neurons, but not lamina II, which was quite strange. I’m not sure if it’s this particular gene (a fluorophore) or the serotype (AAV9) or the injection method (maybe we went too deep?).
I’ve seen AAV1, AAV8 and AAV9 used for this purpose.
AAV2/8 works well to infect neurons in all of the dorsal horn without infecting DRG or brain if you inject directly into the dorsal horn tissue. As you can see from our paper you mentioned Peirs et al Neuron, 2015. In supplemental we show results of several serotypes. See also the paper from Wenqin Luo’s group in Neuron 2016 where they also used AAV8.
AAV9 can easily spread to DRG. You may have gone too deep. I assume you were not using a cre mouse line? hSyn is a good promotor for dorsal horn neurons.
I’ll use AAV8 in the future for DH neurons. I’ve used AAV9 for intrathecal injections, and it’s the best serotype for DRG transduction, in my hands. It did transduce the DH robustly too, but mostly in the deep DH, as I mentioned. It probably also infected DRGs through their terminals in the DH, but for my purposes in this experiment, this wasn’t a prime concern.
I wasn’t using a Cre line. I agree I may have gone too deep, but strangely, I saw lamina I neurons transduced, but in several animals, lamina II was distinctly lacking expression. It could be a serotype issue or a transgene issue. I’ll figure that out when/if I try AAV8.
Hi@achamess, would you like to share your findings about AAV8 and AAV9 in the transduction of DH neurons? We did not see a lot of DH neuronal transduction in the superficial lamina when we used AAV9-Syn-GCAMP. Thank you!
I never put enough time into it evaluate fully. Where are you getting your virus from? I’ve found this to matter. Addgene has reliable virus. Try other transgenes. GCaMP isn’t always the brightest. Try tomato.