Single-particle tracking uncovers dynamics of glutamate-induced retrograde transport of NF-κB p65 in living neurons

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 
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 (Dmean) in neurites from 0.12±0.05 to 0.61±0.03  μm2/s 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 10.9±1.9 to 15±4.9  μm/s, whereas a velocity increase from 9±1.3 to 14±3  μm/s 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.
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Author:Darius Widera, Christin Klenke, Deepak Nair, Meike Heidbreder, Sebastian Malkusch, Jean-Baptiste Sibarita, Daniel Choquet, Barbara Kaltschmidt, Mike Heilemann, Christian Kaltschmidt
URN:urn:nbn:de:hebis:30:3-504810
DOI:http://dx.doi.org/10.1117/1.NPh.3.4.041804
ISSN:2329-4248
ISSN:2329-423X
Pubmed Id:http://www.ncbi.nlm.nih.gov/pubmed?term=27226975
Parent Title (English):Neurophotonics
Publisher:SPIE
Place of publication:Bellingham, Wash.
Document Type:Article
Language:English
Year of Completion:2016
Date of first Publication:2016/05/18
Publishing Institution:Universitätsbibliothek Johann Christian Senckenberg
Release Date:2019/07/15
Tag:NF-kappaB; neurons; retrograde transport; single molecule; single-particle tracking with photoactivated-localization microscopy
Volume:3
Issue:4, Art. 041804
Pagenumber:7
First Page:041804-1
Last Page:041804-6
Note:
© The Authors. Published by SPIE under a Creative Commons Attribution 3.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.
HeBIS PPN:452225132
Institutes:Biochemie und Chemie
Dewey Decimal Classification:540 Chemie und zugeordnete Wissenschaften
Sammlungen:Universitätspublikationen
Licence (German):License LogoCreative Commons - Namensnennung 3.0

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