Special Section on Super-resolution Microscopy of Neural Structure and Function

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

[+] Author Affiliations
Darius Widera

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

Christin Klenke, Christian Kaltschmidt

University of Bielefeld, Cell Biology, Universitätsstr. 25, 33501 Bielefeld, Germany

Deepak Nair, Daniel Choquet

University of Bordeaux, Interdisciplinary Institute for Neuroscience, 146 rue Léo-Saignat, Bordeaux 33077, France

CNRS UMR 5297, 146 rue Léo-Saignat, Bordeaux 33077, France

Meike Heidbreder

Julius-Maximilians-Universität, Department of Biotechnology and Biophysics, Am Hubland, Würzburg 97074, Germany

Sebastian Malkusch, Mike Heilemann

Johann Wolfgang Goethe-University, Institute for Physical and Theoretical Chemistry, Max-von-Laue-Street 7, Frankfurt 60438, Germany

Jean-Baptiste Sibarita

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

Barbara Kaltschmidt

University of Bielefeld, Cell Biology, Universitätsstr. 25, 33501 Bielefeld, Germany

University of Bielefeld, Molecular Neurobiology, Universitätsstr. 25, Bielefeld 33501, Germany

Neurophoton. 3(4), 041804 (May 18, 2016). doi:10.1117/1.NPh.3.4.041804
History: Received October 30, 2015; Accepted April 18, 2016
Text Size: A A A

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 (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.

Figures in this Article
© 2017 Society of Photo-Optical Instrumentation Engineers

Citation

Darius Widera ; Christin Klenke ; Deepak Nair ; Meike Heidbreder ; Sebastian Malkusch, et al.
"Single-particle tracking uncovers dynamics of glutamate-induced retrograde transport of NF-κB p65 in living neurons", Neurophoton. 3(4), 041804 (May 18, 2016). ; http://dx.doi.org/10.1117/1.NPh.3.4.041804


Tables

Access This Article
Sign in or Create a personal account to Buy this article ($20 for members, $25 for non-members).

Some tools below are only available to our subscribers or users with an online account.

Related Content

Customize your page view by dragging & repositioning the boxes below.

Related Book Chapters

Topic Collections

Advertisement
  • Don't have an account?
  • Subscribe to the SPIE Digital Library
  • Create a FREE account to sign up for Digital Library content alerts and gain access to institutional subscriptions remotely.
Access This Article
Sign in or Create a personal account to Buy this article ($20 for members, $25 for non-members).
Access This Proceeding
Sign in or Create a personal account to Buy this article ($15 for members, $18 for non-members).
Access This Chapter

Access to SPIE eBooks is limited to subscribing institutions and is not available as part of a personal subscription. Print or electronic versions of individual SPIE books may be purchased via SPIE.org.