Retrograde gene delivery to hypoglossal motoneurons using adeno-associated virus serotype 9.

Journal Article

Retrograde viral transport (i.e., muscle to motoneuron) enables targeted gene delivery to specific motor pools. Recombinant adeno-associated virus serotype 9 (AAV9) robustly infects motoneurons, but the retrograde transport capabilities of AAV9 have not been systematically evaluated. Accordingly, we evaluated the retrograde transduction efficiency of AAV9 after direct tongue injection in 129SVE mice as well as a mouse model that displays neuromuscular pathology (Gaa(-/-)). Hypoglossal (XII) motoneurons were histologically evaluated 8 weeks after tongue injection with AAV9 encoding green fluorescent protein (GFP) with expression driven by the chicken β-actin promoter (1 × 10(11) vector genomes). On average, GFP expression was detected in 234 ± 43 XII motoneurons 8 weeks after AAV9-GFP tongue injection. In contrast, tongue injection with a highly efficient retrograde anatomical tracer (cholera toxin β subunit, CT-β) resulted in infection of 818 ± 88 XII motoneurons per mouse. The retrograde transduction efficiency of AAV9 was similar between the 129SVE mice and those with neuromuscular disease (Gaa(-/-)). Routine hematoxylin and eosin staining and cluster of differentiation (CD) immunostaining for T cells (CD3) indicated no persistent inflammation within the tongue or XII nucleus after AAV9 injection. Additional experiments indicated no adverse effects of AAV9 on the pattern of breathing. We conclude that AAV9 can retrogradely infect a significant portion of a given motoneuron pool in normal and dystrophic mice, and that its transduction efficiency is approximately 30% of what can be achieved with CT-β.

Full Text

Duke Authors

Cited Authors

  • ElMallah, MK; Falk, DJ; Lane, MA; Conlon, TJ; Lee, K-Z; Shafi, NI; Reier, PJ; Byrne, BJ; Fuller, DD

Published Date

  • April 2012

Published In

Volume / Issue

  • 23 / 2

Start / End Page

  • 148 - 156

PubMed ID

  • 22693957

Electronic International Standard Serial Number (EISSN)

  • 1946-6544

International Standard Serial Number (ISSN)

  • 1946-6536

Digital Object Identifier (DOI)

  • 10.1089/hgtb.2012.009

Language

  • eng