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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.

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.

Abstract

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.

KEYWORDS:

NF-kappaB; neurons; retrograde transport; single molecule; single-particle tracking with photoactivated-localization microscopy

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