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

Widera, D. ORCID: https://orcid.org/0000-0003-1686-130X, Klenke, C., Nair, D., Heidbreder, M., Malkusch, S., Sibarita, J.-B., Choquet, D., Kaltschmidt, B., Heilemann, M. and Kaltschmidt, C. (2016) Single-particle tracking uncovers dynamics of glutamate-induced retrograde transport of NF-κB p65 in living neurons. Neurophotonics, 3 (4). 041804. ISSN 2329-4248 doi: 10.1117/1.NPh.3.4.041804

Abstract/Summary

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 calibre 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. In this study 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 µm2/s 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.

Item Type	Article
URI	https://reading-clone.eprints-hosting.org/id/eprint/63751
Identification Number/DOI	10.1117/1.NPh.3.4.041804
Refereed	Yes
Divisions	Life Sciences > School of Chemistry, Food and Pharmacy > School of Pharmacy > Division of Pharmacology
Publisher	SPIE
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