Biology Reference
In-Depth Information
Fig. 2
(
a
and
b
) reproduced from ref. [
9
]. MNs are incubated with CAdV2-Cy3 for 45 min and imaged by confo-
cal microscopy at 1 frame every 5 s. (
a
) Individual frames are shown. The cell body is located to the
left
.
Arrowheads
show a virion being retrogradely transported,
asterisks
an anterograde virion stopping and chang-
ing to a retrograde direction. (
b
) Kymograph of the corresponding movie with a retrograde CAdV-2 highlighted
in
blue
. The viral particle labeled by the
asterisk
in (
a
) appears in
red
. (
c
) Picture of a microfl uidic chambers.
One set of compartment is fi lled with colorant with a smaller volume than the other compartment. The absence
of diffusion of the dye shows the unidirectional fl ux.
Red oval
highlight the microgrooves. (
d
) Example of trans-
ported CAdV2-Cy3 in axons growing in the microgrooves. Scale bars: 5
μ
m
contains 200
L of medium to ensure a constant fl ow towards
the axons and avoid diffusion of the probes inside the micro-
grooves. Axonal transport is visualized using the same param-
eters than described above.
μ
4
Notes
1. Most of the methods and data generated can be found in ref. [
9
].
2. All labeled-ligands have to be tested for infectivity/functional-
ity to ensure that the labeling procedure did not affect their
structure, which could lead to nonspecifi c cellular uptake.
3. Neuronal cultures described here are MNs and hippocampal
neurons. However, these assays can be applied to virtually any
type of neurons that can be cultured in vitro such as for instance
dorsal root ganglia neurons, sympathetic neurons, or cortical
neurons.
