Biology Reference
In-Depth Information
Chapter 5
Study of Adenovirus and CAR Axonal Transport
in Primary Neurons
Charleine Zussy and Sara Salinas
Abstract
Vectors derived from the canine adenovirus serotype 2 (CAV-2) possess a high neurotropism and effi cient
retrograde transport that lead to widespread neuronal transduction in the central nervous system (CNS) of vari-
ous 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 traffi cking 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 fl uorescently labeled virions or recombinant viral
proteins. Using this approach, we could recapitulate the mechanisms responsible for long-range transport of
adenovirus in neurons.
Key words
Adenovirus, Coxsackievirus and adenovirus receptor, Axonal transport, Fluorescent-
ligands, Live-cell imaging
1
Introduction
Adenoviral-mediated gene transfer requires a clear understanding of
virus-receptor interaction that underlines cell tropism. In this line,
there is now a growing amount of data regarding the intracellular
traffi cking mechanisms involved during vector entry, mainly in epi-
thelial and nonpolarized cells. Indeed, characterizing at the molecu-
lar level the receptor-mediated entry and signaling of viral vectors is
crucial prior their clinical use. Adenoviral vectors have been initially
used for brain cell transduction in the early 1990s [
1
]. In the central
nervous system (CNS), human adenovirus-derived vectors (HAdV)
can infect numerous cell types including neurons, astrocytes, and
oligodendrocytes [
2
]. On the other hand, vectors derived from the
nonhuman AdV CAdV-2 (canine adenovirus type 2) have a high
neurotropism and effi cient axonal retrograde transport in the CNS
of rodents, dogs, and primates that allow a widespread neuronal
transduction. In this light, CAdV vectors represent promising tools
for both understanding and treat neuronal pathologies [
2
,
3
].
