18 May 2016 Single-particle tracking uncovers dynamics of glutamate-induced retrograde transport of NF-κB p65 in living neurons
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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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Darius Widera, Darius Widera, Christin Klenke, Christin Klenke, Deepak Nair, Deepak Nair, Meike Heidbreder, Meike Heidbreder, Sebastian Malkusch, Sebastian Malkusch, Jean-Baptiste Sibarita, Jean-Baptiste Sibarita, Daniel Choquet, Daniel Choquet, Barbara Kaltschmidt, Barbara Kaltschmidt, Mike Heilemann, Mike Heilemann, Christian Kaltschmidt, Christian Kaltschmidt, } "Single-particle tracking uncovers dynamics of glutamate-induced retrograde transport of NF-κB p65 in living neurons," Neurophotonics 3(4), 041804 (18 May 2016). https://doi.org/10.1117/1.NPh.3.4.041804 . Submission:

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