Abstract
Vectors derived from the canine adenovirus serotype 2 (CAV-2) possess a high neurotropism and efficient retrograde transport that lead to widespread neuronal transduction in the central nervous system (CNS) of various animals. These abilities are due to the engagement of virions to the coxsackievirus and adenovirus receptor at the surface of neurons, which is linked to the endocytic and axonal transport machineries. The trafficking of CAV-2 and the coxsackievirus and adenovirus receptor (CAR) can be visualized ex vivo by incubating primary neurons (e.g., motoneurons and hippocampal neurons) with fluorescently labeled virions or recombinant viral proteins. Using this approach, we could recapitulate the mechanisms responsible for long-range transport of adenovirus in neurons.
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References
Le Gal La Salle G, Robert JJ, Berrard S, Ridoux V, Stratford-Perricaudet LD, Perricaudet M et al (1993) An adenovirus vector for gene transfer into neurons and glia in the brain. Science 259:988–990
Kremer EJ (2005) Gene transfer to the central nervous system: current state of the art of the viral vectors. Curr Genomics 6:13–39
Bru T, Salinas S, Kremer EJ (2010) An update on canine adenovirus type 2 and its vectors. Viruses 2:2134–2153
Coyne CB, Bergelson JM (2005) CAR: a virus receptor within the tight junction. Adv Drug Deliv Rev 57:869–882
Soudais C, Boutin S, Hong SS, Chillon M, Danos O, Bergelson JM et al (2000) Canine adenovirus type 2 attachment and internalization: coxsackievirus-adenovirus receptor, alternative receptors, and an RGD-independent pathway. J Virol 74:10639–10649
Honda T, Saitoh H, Masuko M, Katagiri-Abe T, Tominaga K, Kozakai I et al (2000) The coxsackievirus-adenovirus receptor protein as a cell adhesion molecule in the developing mouse brain. Mol Brain Res 77:19–28
Patzke C, Max KE, Behlke J, Schreiber J, Schmidt H, Dorner AA et al (2010) The coxsackievirus-adenovirus receptor reveals complex homophilic and heterophilic interactions on neural cells. J Neurosci 30:2897–2910
Henaff D, Salinas S, Kremer EJ (2011) An adenovirus traffic update: from receptor engagement to the nuclear pore. Future Microbiol 6:179–192
Salinas S, Bilsland LG, Henaff D, Weston AE, Keriel A, Schiavo G et al (2009) CAR-associated vesicular transport of an adenovirus in motor neuron axons. PLoS Pathog 5:e1000442
Salinas S, Schiavo G, Kremer EJ (2010) A hitchhiker’s guide to the nervous system: the complex journey of viruses and toxins. Nat Rev Microbiol 8:645–655
Taylor AM, Rhee SW, Jeon NL (2006) Microfluidic chambers for cell migration and neuroscience research. Methods Mol Biol 321:167–177
Taylor AM, Blurton-Jones M, Rhee SW, Cribbs DH, Cotman CW, Jeon NL (2005) A microfluidic culture platform for CNS axonal injury, regeneration and transport. Nat Methods 2:599–605
Kremer EJ, Boutin S, Chillon M, Danos O (2000) Canine adenovirus vectors: an alternative for adenovirus-mediated gene transfer. J Virol 74:505–512
Schoehn G, El Bakkouri M, Fabry CM, Billet O, Estrozi LF, Le L et al (2008) Three-dimensional structure of canine adenovirus serotype 2 capsid. J Virol 82:3192–3203
Deinhardt K, Salinas S, Verastegui C, Watson R, Worth D, Hanrahan S et al (2006) Rab5 and Rab7 control endocytic sorting along the axonal retrograde transport pathway. Neuron 52:293–305
Bohnert S, Schiavo G (2005) Tetanus toxin is transported in a novel neuronal compartment characterized by a specialized pH regulation. J Biol Chem 280:42336–42344
Acknowledgments
We thank Eric Kremer and members of the EKL group, G. Schiavo, and G. Menendez for the microfluidic chambers. S.S. is an Institut National de la Santé et de la Recherche Médicale (INSERM fellow). Work in the laboratory is funded by the European Community’s 7th Framework Programme (FP7/2007–2013; grant 222992—BrainCAV), the French Agence National de la Recherche, E-Rare, the Fondation de France and the Association Française contre les Myopathies.
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Zussy, C., Salinas, S. (2014). Study of Adenovirus and CAR Axonal Transport in Primary Neurons. In: Chillón, M., Bosch, A. (eds) Adenovirus. Methods in Molecular Biology, vol 1089. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-679-5_5
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DOI: https://doi.org/10.1007/978-1-62703-679-5_5
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