Spira Laboratory
home
Lab members
Images
movies
publications
links
suplement movies to research proposal
suplement movies to papers
research interests
contact
Aplysia neuron after axotomy

Supplemental videos for Journal of Cell Biology (JCB)

These files are in AVI format. 

Video 1, 2 and 3. The videos illustrate the phenomena depicted by figure 1. Movie 1 demonstrates EB3-GFP, "comet tails" in the axon. The movie contains 29 images, taken at intervals of 12 seconds. Movie 2 demonstrates the non-polar orientation of microtubules in the soma and the typical MTOC usually observed at the axon hillock as determined by EB3-GFP.  The movie is composed of 70 images, taken at intervals of 4 seconds. Movie 3 illustrates the transport of SR101 labeled endocytotic vesicles. The movie comprises 20 images, taken at intervals of 7.2 seconds. The frames are shown at a rate of 10/s. Scalebars: 10 Ám.

Video 1 (download time approximately 60 seconds)

Video 2 (download time approximately 60 seconds)

Video 3 (download time approximately 60 seconds)

Video 4 and 5. Anterograde transport of EYFP-SNAP-25 labeled vesicles and their accumulation after axotomy. The cell body (unpublished data) is on the left hand side of the screen. Video 4- before and during axotomy. Note thVideo 6. The retrograde transport of SR101 labeled vesicles. This video was taken 75 minutes after washing the SR101 from the bathing solution. The vesicles are retrogradely transported towards the cell body, which is located on the left side of the screen (unpublished data). The video is composed of 60 images, taken at intervals of 5 seconds. The frames are shown at a rate of 10/s.   f 10/s.   

Video 4 (download time approximately 60 seconds)

Video 5 (download time approximately 60 seconds)

Video 6. The retrograde transport of SR101 labeled vesicles. This video was taken 75 minutes after washing the SR101 from the bathing solution. The vesicles are retrogradely transported towards the cell body, which is located on the left side of the screen (unpublished data). The video is composed of 60 images, taken at intervals of 5 seconds. The frames are shown at a rate of 10/s. 

Video 6 (download time approximately 60 seconds)   

Video 7 and 8. The formation of a plus-end trap depends on anterograde transport of components from the cell body to the axon. Axotomy of an intact neuron leads to the formation of a plus end trap (EB3-GFP, green), which concentrates anterogradely-transported vesicles (cherry-SNAP-25, red) as shown by Video 7 (corresponding to Fig. 4 B). The video is composed of 20 images, taken at intervals of 23 seconds. The frames are shown at a rate of 7/s. In contrast, axotomy of an isolated axonal segment (Video 8, corresponds to Fig. 4 C ) does not lead to reorientation of the MTs (EB3-GFP, green) and to accumulation of SNAP-25 (cherry-SNAP-25, red). Video 8 is composed of 15 images, taken at intervals of 7 seconds. The frames are shown at a rate of 10/s.

Video 7 (download time approximately 60 seconds)

Video 8 (download time approximately 60 seconds)

Video 9 and 10. The formation of the MT-based plus-end and minus-end traps cannot be correlated with retrogradely transported retrieved membrane. Pinocytotic vesicles were labeled by SR101 as described for the experiment in figure 5. Axotomy of an intact neuron leads to the formation of a minus end trap in which retrogradely transported vesicles accumulate (Video 9, SR101-red, EB3-GFP-green, corresponds to Fig. 5 A ). The video is composed of 60 images, taken at intervals of 7 seconds. The frames are shown at a rate of 10/s. In contrast, axotomy of an isolated axonal segment does not lead to reorientation of the MTs, although accumulation of SR101 at the distal segment of the axon is detected (Video 10 corresponds to Fig. 5 B). The video is composed of 60 images, taken at intervals of 7 seconds. The frames are shown at a rate of 10/s. 

Video 9 (download time approximately 60 seconds)

Video 10 (download time approximately 60 seconds)

Video 11. The formation of the MT-based plus- and minus-end traps does not depend on the supply of  Golgi derived anterogradely transported vesicles. An EB3-GFP (green) expressing neuron was bathed in 10Ál/ml BFA for 17 hours leading to disruption of the Golgi system. 4 hours before axotomy the cell was injected with  SNAP-25-cherry mRNA. Following axotomy the MTs formed the plus and minus traps. The cherry -SNAP25 proteins (red) was evenly distributed throughout the axoplasm and did not accumulate within the GCOC. The upper and lower movies were taken 1 and 11 minutes after axotomy, respectively. The upper movie is composed of 60 images and the lower one of 30 images. The images were taken at intervals of 6 seconds. The frames are shown at a rate of 10/s.

Video 11- up (download time approximately 60 seconds) 

Video 11- Down (download time approximately 60 seconds)

 

                                                    This site was originated by Prof. Micha E. Spira. We welcome your questions, suggestions and comments. 
                                                    Copyright ©, 1997, The Hebrew University of Jerusalem. All Right Reserved.