rAAV transduction in the myenteric and submucous plexus of mouse ileum and colon

      Background: Recombinant adeno-associated viruses (rAAV) are a widely used type of viral transduction vectors. Although the efficiency of this transduction method has been extensively studied in the (central) nervous system, little is known about the targeting of the enteric nervous system (ENS) by rAAV. Our aim was hence to assess the transduction in the ENS by rAAV2/8 and rAAV2/9. Methods: Mice were injected i.v. with rAAV at postnatal day 1. The rAAV vectors encoded the transgene for enhanced green fluorescent protein (eGFP) under the control of a CMV promoter. Using immunohistochemical staining for neurochemical subclasses of enteric neurons in whole mount preparations of colon and ileum, eGFP expression in the ENS was evaluated at postnatal day 35. Results: Both rAAV2/8 and rAAV2/9 transduction resulted in eGFP expression in a quarter of the myenteric and ileal submucous neurons, whereas eGFP expression was remarkably lower in the submucous plexus of the colon. eGFP-labeled enteric neurons could be labeled with VIP, calretinin, calbindin or neuronal NOS, indicating that all functional enteric neuronal subtypes are susceptible to rAAV transduction. Staining with S100 or GFAP did not reveal any significant transduction of enteric glia. Conclusion: Our results on rAAV-transduction of myenteric neurons corroborate the recent findings of Gombash et al. (Front Mol Neurosci 2014) and expand the application of rAAV transduction to the submucous plexus. This approach allows manipulation of specific neuronal subpopulations in vivo, which is a useful research tool in the short term and a possible therapeutic strategy in the longer term. This study is funded by grant G019314N of the Research Foundation – Flanders (FWO).
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