Single-particle tracking uncovers dynamics of glutamate-induced retrograde transport of NF-κB p65 in living neurons.
Neurophoton. 2016-05-18; 3(4): 041804
DOI: 10.1117/1.nph.3.4.041804
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1. Neurophotonics. 2016 Oct;3(4):041804. doi: 10.1117/1.NPh.3.4.041804. Epub 2016
May 18.
Single-particle tracking uncovers dynamics of glutamate-induced retrograde
transport of NF-κB p65 in living neurons.
Widera D(1), Klenke C(2), Nair D(3), Heidbreder M(4), Malkusch S(5), Sibarita
JB(6), Choquet D(3), Kaltschmidt B(7), Heilemann M(5), Kaltschmidt C(2).
Author information:
(1)University of Bielefeld, Cell Biology, Universitätsstr. 25, 33501 Bielefeld,
Germany; University of Reading, School of Pharmacy, Stem Cell Biology and
Regenerative Medicine, Whiteknights, Reading RG6 6UB, United Kingdom.
(2)University of Bielefeld , Cell Biology, Universitätsstr. 25, 33501 Bielefeld,
Germany.
(3)University of Bordeaux, Interdisciplinary Institute for Neuroscience, 146 rue
Léo-Saignat, Bordeaux 33077, France; CNRS UMR 5297, 146 rue Léo-Saignat, Bordeaux
33077, France.
(4)Julius-Maximilians-Universität , Department of Biotechnology and Biophysics,
Am Hubland, Würzburg 97074, Germany.
(5)Johann Wolfgang Goethe-University , Institute for Physical and Theoretical
Chemistry, Max-von-Laue-Street 7, Frankfurt 60438, Germany.
(6)University of Bordeaux, Interdisciplinary Institute for Neuroscience, 146 rue
Léo-Saignat, Bordeaux 33077, France; CNRS UMR 5297, 146 rue Léo-Saignat, Bordeaux
33077, France; Bordeaux Imaging Center, UMS 3420 CNRS, US4 INSERM, France.
(7)University of Bielefeld, Cell Biology, Universitätsstr. 25, 33501 Bielefeld,
Germany; University of Bielefeld, Molecular Neurobiology, Universitätsstr. 25,
Bielefeld 33501, Germany.
Retrograde transport of NF-κB from the synapse to the nucleus in neurons is
mediated by the dynein/dynactin motor complex and can be triggered by synaptic
activation. The caliber of axons is highly variable ranging down to 100 nm,
aggravating the investigation of transport processes in neurites of living
neurons using conventional light microscopy. We quantified for the first time the
transport of the NF-κB subunit p65 using high-density single-particle tracking in
combination with photoactivatable fluorescent proteins in living mouse
hippocampal neurons. We detected an increase of the mean diffusion coefficient
([Formula: see text]) in neurites from [Formula: see text] to [Formula: see text]
after stimulation with glutamate. We further observed that the relative amount of
retrogradely transported p65 molecules is increased after stimulation. Glutamate
treatment resulted in an increase of the mean retrograde velocity from [Formula:
see text] to [Formula: see text], whereas a velocity increase from [Formula: see
text] to [Formula: see text] was observed for anterogradely transported p65. This
study demonstrates for the first time that glutamate stimulation leads to an
increased mobility of single NF-κB p65 molecules in neurites of living
hippocampal neurons.
DOI: 10.1117/1.NPh.3.4.041804
PMCID: PMC4870386
PMID: 27226975