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Videos

Gunther Gerisch's Demonstration Movies:

1.  Micropipet-mediated aggregation MPG

2.  Single Dictyostelium cell exposed to a cyclic AMP gradient from a micropipette.  Concentration of cyclic AMP changes ~20% across the cell. MPG

3. Stepwise chemotactic movements of Dictyostelium cells towards an aggregation center. MPG

4.  Chemotaxis of neutrophil chasing a bacteriummov.gif (1427 bytes)

GFP-Tagged Chemoattractant Receptors and G-protein Subunits in Chemotaxing D. discoideum Cells

5.  Green fluorescent protein-tagged receptors in living cells undergoing chemotaxis.  Micropipette is initially at the top of the frame, and then moved to the lower portion of the frame.  avi.gif (1407 bytes) mov.gif (1427 bytes)

6.  Green fluorescent protein-tagged receptors in living cells undergoing chemotaxis.  Micropipette was moved to the upper right hand portion of the frame. avi.gif (1407 bytes) mov.gif (1427 bytes)

7.  Colorized confocal image of GFP-Gb in chemotaxing cells.  Gb-GFP was used to rescue the null phenotype of Gb null cells.  avi.gif (1407 bytes)
 

PH-GFP Biosensors Reporting PIP3 Levels in Chemotaxing D. discoideum Cells

8.  Kinetic analysis of  translocation of CRAC-GFP in response to a uniform increase in chemoattractant concentration. Frames are taken every two seconds and chemoattractant was added just before cell goes out of focus. avi.gif (1407 bytes)  

9.  CRAC-GFP translocates to the leading edge of newly elicited pseudopods in chemotactically moving cells. The micropipette is located in the center of the converging cells. avi.gif (1407 bytes)  

10.  Wild-type cells expressing PH-GFP carrying out chemotaxis towards cyclic AMP. avi.gif (1407 bytes)

11.  Same cells as 9 treated with LY429033. avi.gif (1407 bytes)

12.  Dynamics of the CRAC-GFP redistribution. A large increment in chemoattractant was applied by increasing the microinjector pressure and the cells immediately respond. As the gradient of cAMP is restored, the polarization of CRAC-GFP is re-established. avi.gif (1407 bytes)  

13.  Response of CRAC-GFP to an approaching wave of chemoattractant. Pipette located on the right side of the cell was briefly "turned on" for six seconds at three times during the sequence.  avi.gif (1407 bytes)

14.  Response of CRAC-GFP to an approaching wave of chemoattractant. Pipette located on the left side of the same cell as in #6 was briefly "turned on" for six seconds at three times during the sequence. avi.gif (1407 bytes)  

15.  Translocation of CRAC-GFP in cells lacking actin filament formation. Cells chemotaxing towards pipette in lower portion of field were poisoned with Latrunculin A and followed for the next several minutes. avi.gif (1407 bytes)  

16.  Latrunculin immobilized cells were stimulated with a uniform increment in cyclic AMP.avi.gif (1407 bytes)

17.  Latrunculin immobilized cells exposed to micropipette containing cyclic AMP.avi.gif (1407 bytes)

18.  Latrunculin A immobilized cells were exposed to a pipette in upper portion of field.  The pipet was then moved around the cells and the binding sites for PH-domains move around the cell on the inner face of the membrane. avi.gif (1407 bytes) 

19.  Polarized translocation of PH-GFP in highly polarized cells.   Micropipet was located in the lower region beyond the field.  Cells were rapidly exposed to a  saturating dose of attractant. avi.gif (1407 bytes)

20.  Latrunculin immobilized cells were exposed to a micropipette containing cyclic AMP.  Note the directional responses of the cell as the micropipette is repositioned. avi.gif (1407 bytes)

21.  Latrunculin immobilized cell was exposed to two opposing cyclic AMP gradients (see two micropipettes).  The strength of the two gradients was alternated during the video. avi.gif (1407 bytes)

22.  Cells expressing PH-GFP carrying out phagocytosis of yeast particles.  Total time of video is ~4 minutes. avi.gif (1407 bytes)

Single Molecule Imaging by Masahiro Ueda and Toshio Yanagida

23.  Single molecule imaging of the binding of CY3-cyclic AMP to the basal surface of a cell.  Lifetime of binding events is ~1.5 seconds.avi.gif (1407 bytes)

24.  Single molecule imaging of PH-GFP molecules moving from the cytosol to the plasma membrane on the basal surface of the cell. avi.gif (1407 bytes)

Role of PTEN in Directional Sensing

25.  Pten- cells expressing PH-GFP.  Cells were exposed to a sudden increase in cyclic AMP (white flash).  Total time of video represents ~3.5 minutes of stimulation. avi.gif (1407 bytes)

26.   Chemotaxis of wild-type cells expressing PH-GFP towards cyclic AMP filled micropipette in lower left hand corner of field.  avi.gif (1407 bytes)

27.  Chemotaxis of pten- cells towards cyclic AMP filled micropipette in upper right hand corner of field.  Cells are expressing PH-GFP. avi.gif (1407 bytes)

28.  Distribution of PTEN-GFP in cells chemotaxing towards cyclic AMP.  Note that PTEN localizes to the back of the cell. avi.gif (1407 bytes)

29. Pten- cells expressing PH-GFP exposed to a cyclic AMP gradient coming from a micropipette in the middle of the field. avi.gif (1407 bytes)

30.  Same cells as in 27 treated with 30 μm mg of LY429033.  avi.gif (1407 bytes)

Kees Wiejer and Florian Seigert's Videos

Rex  Chisholm's Videos

 

This page was last edited 03/25/2005