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

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

单粒子追踪揭示了活体神经元中谷氨酸诱导的 NF-κB p65 逆行运输的动态

DOI:
10.1117/1.nph.3.4.041804
复制
发表时间:
2016
期刊:
影响因子:
5.3
通讯作者:
Kaltschmidt C
Kaltschmidt C
中科院分区:
医学2区
文献类型:
--
作者:
Widera D;Klenke C;Nair D;Heidbreder M;Malkusch S;Sibarita JB;Choquet D;Kaltschmidt B;Heilemann M;Kaltschmidt C

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在神经元中,NF-κB从突触向细胞核的逆行转运是由动力蛋白/动力蛋白运动复合物介导的,并可由突触激活触发。轴突的直径变化很大,可低至100纳米,这使得使用常规光学显微镜研究活神经元的神经突运输过程变得更加困难。我们首次使用高密度单颗粒跟踪结合光激活荧光蛋白定量了NF-κB亚基p65在活体小鼠海马神经元中的转运。我们检测到谷氨酸刺激后神经突的平均扩散系数()增加。我们进一步观察到,刺激后p65分子逆行转运的相对数量增加。谷氨酸处理导致p65的平均逆行速度增加,而p65的顺行速度增加。本研究首次证明了谷氨酸刺激可导致活海马神经元神经突中单个NF-κB p65分子的移动性增加。
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 () in neurites fromtoafter 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 fromto, whereas a velocity increase fromtowas 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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