The Journal of Neuroscience, January 1, 2003, ():
KIF17 Dynamics and Regulation of NR2B Trafficking in Hippocampal Neurons
J. Neurosci.
Guillaud et al. 23 (1): 131.
Supplemental Data
Figure 1: Quantification of the overexpression levels of YFP-KIF17 in hippocampal neurons. After 10 days of culture, hippocampal neurons were transfected with YFP-KIF17, fixed and immunostained with KIF17 pab. Fluorescence intensity was measured using the fluorescence profile function of LSM510 software in three different regions of the dendrite (proximal, middle and distal) and in the cell body of transfected or non-transfected neurons. After overexpression of YFP-KIF17 the KIF17 expression level increased by 20 to 30 %.
Files in this Data Supplement:
- Figure 1
- Video 1
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Fluorescence recovery after photobleaching of YFP-KIF17 in the proximal dendrite of 10 days in vitro (d.i.v.) hippocampal neuron. After 10 days of culture, hippocampal neurons were transfected with YFP-KIF17 and directly observed under confocal microscope. A 15-µm area on the apical dendrite was bleached and the fluorescence recovery was monitored every 5 seconds. Recovery was achieved from both the proximal part and the distal part of the bleached area. Small vesicular structures moved within the area and fluorescence was completely restored after 20 seconds
- Video 2
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Dynamics of YFP-KIF17 single vesicles in the distal dendrite of 10 d.i.v. hippocampal neuron. After 10 days of culture, hippocampal neurons were transfected with YFP-KIF17 and directly observed under confocal microscope. Images were acquired every second and movement was monitored for 30 seconds. Several vesicles are moving anterogradely toward the tip of the dendrite while some larger structures remain immobile.
- Video 3
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3D reconstruction and animation of YFP-KIF17 vesicles in dendrite of 10 d.i.v. hippocampal neuron. After 10 days of culture, hippocampal neurons were transfected with YFP-KIF17, fixed and processed for immunodetection of synaptic clusters using anti-PSD95 mAb. Rendering of 3D images from confocal Z stacks files was performed using Autodeblur and Autovisualize. YFP-KIF17 vesicles (green) are restricted only to the shaft of the dendrite and are excluded from the postsynaptic region as shown with PSD95 staining (blue).
