The Journal of Neuroscience, April 1, 2003, ():
Visualization of Microtubule Growth in Cultured Neurons via the Use of EB3-GFP (End-Binding Protein 3-Green Fluorescent Protein)
J. Neurosci.
Stepanova et al. 23 (7): 2655.
Movies
Files in this Data Supplement:
- Movie 2
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EB3-GFP movement in transfected COS-1 cells.
COS-1 cells, transfected with EB3-GFP, were analyzed 48 hours after transfection by confocal microscopy. Cells were maintained in normal culture medium at 37 0C. Shown is an area of 552 x 586 pixels (78 by 83 micrometers), in which 1 transfected cell is completely visible and part of another cell is also shown to the right. A total of 100 images (0.9 second per image) were collected every 3 seconds. Notice that the EB3-GFP signal at microtubule plus ends, shown here, is brighter than the GFP-CLIP115 signal shown in movie 1. See legend to movie 1 for further details.
- Movie 3
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EB3-GFP movement in live transfected neurons.
Embryonic mouse hippocampal neurons were electroporated with EB3-GFP encoding DNA and cells expressing low levels of the fusion protein were examined by confocal microscopy. Cells were maintained at 37 0C in conditioned N2 culture medium. The argon laser was set at 1-2 % to minimize bleaching effects. This movie shows EB3-GFP movement in a complete neuron (area: 1024 x 1024 pixels, or 114 x 114 micrometers). Notice that part of the axon is covered by a non-transfected cell. Plus end movement occurs in all neuronal compartments. A total of 100 images were acquired every 1.5 seconds.
- Movie 5
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EB3-GFP movement in infected PC cultures.
See movie 4 for details. A total of 100 images were acquired every 2 seconds (area: 512 x 512 pixels, or 29 x 29 micrometers). Notice the bidirectional movements of EB3-GFP dashes.
- Movie 6
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EB3-GFP movement in infected hippocampal cultures.
Embryonic mouse hippocampal neurons of 10 DIV were infected with SFV particles, containing EB3-GFP encoding RNA. Cells were maintained at 37 0C in conditioned culture medium. Cells expressing low levels of the fusion protein were examined 5-8 hours post-infection, by confocal microscopy. In this movie EB3-GFP movement is visualized in an axon. A total of 100 images were acquired every 2 seconds (area: 784 x 246 pixels; or 117 x 36 micrometer). Notice that the vast majority of EB3-GFP dashes moves unidirectionally.
- Movie 7
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EB3-GFP movement in infected hippocampal cultures.
See movie 6 for details. In this movie GFP-EB3 movements in a retracting axon are visualized. A total of 100 images were acquired every 2 seconds (area: 268 x 512 pixels; or 27 x 52 micrometers). Notice how, in spite of the fact that this growth cone is retracting, EB3-GFP dashes continue to penetrate the growth cone area.
