# Neuroimmune–Glia Interactions in the Sensory Ganglia Account for the Development of Acute Herpetic Neuralgia **URL:** \ **Category:** Journal Club\ **Created:** [September 30, 2017, 7:41pm UTC](https://forum.painresearcher.net/t/neuroimmune-glia-interactions-in-the-sensory-ganglia-account-for-the-development-of-acute-herpetic-neuralgia/283 "2017-09-30T19:41:56Z")\ **Posts on this page:** 12\ **Page:** 1
**Author:** ![achamess](https://yyz2.discourse-cdn.com/flex030/user_avatar/forum.painresearcher.net/achamess/32/534_2.png) [@achamess](https://forum.painresearcher.net/u/achamess)\ **Post date:** [September 30, 2017, 7:41pm UTC](https://forum.painresearcher.net/t/neuroimmune-glia-interactions-in-the-sensory-ganglia-account-for-the-development-of-acute-herpetic-neuralgia/283/1 "2017-09-30T19:41:56Z")
Very nice paper from @thicunha > **[Neuroimmune–Glia Interactions in the Sensory Ganglia Account for the...](https://www.jneurosci.org/content/37/27/6408)** > > Herpetic neuralgia is the most important symptom of herpes zoster disease, which is caused by Varicella zoster . Nevertheless, the pathophysiological mechanisms involved in herpetic neuralgia are not totally elucidated. Here, we examined the... Neuroimmune–Glia Interactions in the Sensory Ganglia Account for the Development… @thicunha there are a lot of interesting methodology things here. For the shRNA, were those plasmids yiu transfected? And for the HSV infection, you put the virus right on the infected skin rather than injection? Was this superior to injection? ---
**Author:** ![achamess](https://yyz2.discourse-cdn.com/flex030/user_avatar/forum.painresearcher.net/achamess/32/534_2.png) [@achamess](https://forum.painresearcher.net/u/achamess)\ **Post date:** [October 2, 2017, 7:09pm UTC](https://forum.painresearcher.net/t/neuroimmune-glia-interactions-in-the-sensory-ganglia-account-for-the-development-of-acute-herpetic-neuralgia/283/2 "2017-10-02T19:09:10Z")
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**Author:** ![achamess](https://yyz2.discourse-cdn.com/flex030/user_avatar/forum.painresearcher.net/achamess/32/534_2.png) [@achamess](https://forum.painresearcher.net/u/achamess)\ **Post date:** [October 2, 2017, 7:09pm UTC](https://forum.painresearcher.net/t/neuroimmune-glia-interactions-in-the-sensory-ganglia-account-for-the-development-of-acute-herpetic-neuralgia/283/3 "2017-10-02T19:09:46Z")
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**Author:** ![thicunha](https://yyz2.discourse-cdn.com/flex030/user_avatar/forum.painresearcher.net/thicunha/32/386_2.png) [@thicunha](https://forum.painresearcher.net/u/thicunha)\ **Post date:** [October 3, 2017, 7:17pm UTC](https://forum.painresearcher.net/t/neuroimmune-glia-interactions-in-the-sensory-ganglia-account-for-the-development-of-acute-herpetic-neuralgia/283/4 "2017-10-03T19:17:01Z")
Dear @achamess Alex, thank you for the comments on our paper. Regarding your questions, we have used in vivo-jetPEI® for transfection ([http://www.polyplus-transfection.com/products/in-vivo-jetpei/](http://www.polyplus-transfection.com/products/in-vivo-jetpei/)), the first transfected reagents approved for human use. We confirmed the transfection by following the GFP expression, which is part of our plasmids. Regarding the infection, we did a scarification in the region before to apply a solution containing the virus (Takasaki I, Andoh T, Shiraki K, Kuraishi Y (2000a) Allodynia and hyperalgesia induced by herpes simplex virus type-1 infection in mice. Pain 86:95–101). We already tried the injection into the hindpaw and the mouse also developed mechanical hypersensitivity but we did not see any skin lesion. ---
**Author:** ![achamess](https://yyz2.discourse-cdn.com/flex030/user_avatar/forum.painresearcher.net/achamess/32/534_2.png) [@achamess](https://forum.painresearcher.net/u/achamess)\ **Post date:** [October 3, 2017, 8:33pm UTC](https://forum.painresearcher.net/t/neuroimmune-glia-interactions-in-the-sensory-ganglia-account-for-the-development-of-acute-herpetic-neuralgia/283/5 "2017-10-03T20:33:58Z")
Thanks @thicunha. Did you see any expression in spinal neurons? I’m interested in using invivo-JetPEI for getting siRNA into spinal neurons. ---
**Author:** ![thicunha](https://yyz2.discourse-cdn.com/flex030/user_avatar/forum.painresearcher.net/thicunha/32/386_2.png) [@thicunha](https://forum.painresearcher.net/u/thicunha)\ **Post date:** [October 4, 2017, 12:19pm UTC](https://forum.painresearcher.net/t/neuroimmune-glia-interactions-in-the-sensory-ganglia-account-for-the-development-of-acute-herpetic-neuralgia/283/6 "2017-10-04T12:19:43Z")
@achamess, We detected the expression of GFP in spinal cord tissue, unfortunately, we only checked using PCR. I guess spinal neurons are transfected, but we need to check. Have you tried lentivirus or AAV ([http://blog.addgene.org/aavs-created-for-gene-delivery-to-the-cns-and-pns](http://blog.addgene.org/aavs-created-for-gene-delivery-to-the-cns-and-pns)). This AVV seems to be nice. ---
**Author:** ![achamess](https://yyz2.discourse-cdn.com/flex030/user_avatar/forum.painresearcher.net/achamess/32/534_2.png) [@achamess](https://forum.painresearcher.net/u/achamess)\ **Post date:** [October 4, 2017, 2:17pm UTC](https://forum.painresearcher.net/t/neuroimmune-glia-interactions-in-the-sensory-ganglia-account-for-the-development-of-acute-herpetic-neuralgia/283/7 "2017-10-04T14:17:21Z")
Thanks @thicunha. AAV works well and I use it when I can. But I don’t have an AAV with an shRNA to the gene I’m targeting. So I’m restricted to using siRNA. Intrathecal is much easier and faster than intraspinal viral injections. I’ve seen people do this before and your recent results with the plasmid and JetPEI make me think siRNA will work pretty well too. ---
**Author:** ![thicunha](https://yyz2.discourse-cdn.com/flex030/user_avatar/forum.painresearcher.net/thicunha/32/386_2.png) [@thicunha](https://forum.painresearcher.net/u/thicunha)\ **Post date:** [October 4, 2017, 2:30pm UTC](https://forum.painresearcher.net/t/neuroimmune-glia-interactions-in-the-sensory-ganglia-account-for-the-development-of-acute-herpetic-neuralgia/283/8 "2017-10-04T14:30:44Z")
@achamess Perfect. Please let me know if you have success with Jetpei and siRNA ---
**Author:** ![tberta](https://yyz2.discourse-cdn.com/flex030/user_avatar/forum.painresearcher.net/tberta/32/145_2.png) [@tberta](https://forum.painresearcher.net/u/tberta)\ **Post date:** [October 5, 2017, 7:09pm UTC](https://forum.painresearcher.net/t/neuroimmune-glia-interactions-in-the-sensory-ganglia-account-for-the-development-of-acute-herpetic-neuralgia/283/9 "2017-10-05T19:09:35Z")
We are currently using i.t. injections of JetPEI with siRNA to target DRG neurons. Our test of a single injection with a fluorescent siRNA showed the transfection of only DRG neurons (~40-70%) and no cells in the spinal cord! Alex: You may need multiple injections to also target spinal cord cells or use other transfection agents. We tested various agents in our lab and most of them work great in vitro, but very poorly in vivo - sigh! ---
**Author:** ![thicunha](https://yyz2.discourse-cdn.com/flex030/user_avatar/forum.painresearcher.net/thicunha/32/386_2.png) [@thicunha](https://forum.painresearcher.net/u/thicunha)\ **Post date:** [October 5, 2017, 8:28pm UTC](https://forum.painresearcher.net/t/neuroimmune-glia-interactions-in-the-sensory-ganglia-account-for-the-development-of-acute-herpetic-neuralgia/283/10 "2017-10-05T20:28:51Z")
@achamess I have look again in our data and @tberta is right. Whereas we have 200X increase (compared with the background) in GFP expression in the DRGs we only see 3X in the spinal cord. Another point, we have injected (it) 5 consecutive days. ---
**Author:** ![achamess](https://yyz2.discourse-cdn.com/flex030/user_avatar/forum.painresearcher.net/achamess/32/534_2.png) [@achamess](https://forum.painresearcher.net/u/achamess)\ **Post date:** [October 5, 2017, 8:34pm UTC](https://forum.painresearcher.net/t/neuroimmune-glia-interactions-in-the-sensory-ganglia-account-for-the-development-of-acute-herpetic-neuralgia/283/11 "2017-10-05T20:34:59Z")
Thanks @thicunha and @tberta. This helps me out a lot. I might not waste my time. @thicunha: In your case, you used a plasmid containing shRNA vs naked siRNA. @tberta: In your case, you did a single siRNA injection. This protocol from Rohini Kuner’s lab (who always does excellent work, IMO), suggests siRNAs can knockdown spinal cell gene expression (it’s modest though). I may still try multi-day naked siRNA, but your experiences diminish my enthusiasm. I’ll let you know how it goes. Thank you for sharing so openly. This kind of discussion is exactly why this Pain Researcher site was created! > > > This report describes a step-by-step guide to the technique of acute intrathecal needle injections in a noninvasive manner, i.e. independent of catheter implantation. The technical limitation of this surgical technique lies in the finesse of the hands. The injection is rapid, especially for a trained experimenter, and since tissue disruption with this technique is minimal, repeated injections are possible; moreover immune reaction to foreign tools (e.g. catheter) does not occur, thereby giving a better and more specific read out of spinal cord modulation. Since the application of the substance is largely limited to the target region of the spinal cord, drugs do not need to be applied in large dosages, and more importantly unwanted effects on other tissue, as observed with a systemic delivery, could be circumvented(1,2). Moreover, we combine this technique with in vivo transfection of nucleic acid with the help of polyethylenimine (PEI) reagent(3), which provides tremendous versatility for studying spinal functions via delivery of pharmacological agents as well as gene, RNA, and protein modulators. ---
**Author:** ![thicunha](https://yyz2.discourse-cdn.com/flex030/user_avatar/forum.painresearcher.net/thicunha/32/386_2.png) [@thicunha](https://forum.painresearcher.net/u/thicunha)\ **Post date:** [October 5, 2017, 8:39pm UTC](https://forum.painresearcher.net/t/neuroimmune-glia-interactions-in-the-sensory-ganglia-account-for-the-development-of-acute-herpetic-neuralgia/283/12 "2017-10-05T20:39:36Z")
@achamess In our case we have used a plasmid containing shRNA vs naked shRNA. Thank you for the excellent initiative to create this forum. I’m going to ask all my students to became members and follow these discussions.