Open Access Highly Accessed Research article

Curvature recognition and force generation in phagocytosis

Margaret Clarke1, Ulrike Engel2, Jennifer Giorgione1, Annette Müller-Taubenberger3, Jana Prassler4, Douwe Veltman5 and Günther Gerisch4*

Author Affiliations

1 Program in Genetic Models of Disease, Oklahoma Medical Research Foundation, Oklahoma City, OK 73121, USA

2 Nikon Imaging Center at the University of Heidelberg, Bioquant, 69120 Heidelberg, Germany

3 Institut für Zellbiologie, Ludwig-Maximilians-Universität München, 80336 München, Germany

4 Max-Planck-Institut für Biochemie, 82152 Martinsried, Germany

5 Beatson Institute for Cancer Research, Glasgow G61 1BD, UK

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BMC Biology 2010, 8:154  doi:10.1186/1741-7007-8-154

Published: 29 December 2010

Additional files

Additional file 1:

Movie 1 - Attack of two budded yeast by a cell expressing LimEΔ-GFP as a label for filamentous actin. The fluorescence is superimposed in green on greyscale brightfield images. Protrusions surrounding the particle are fluctuating between forth and back movement. This movie shows the same events as Figure 1. Frame to frame interval is 4 seconds. All Additional files are movies showing phagocytes of Dictyostelium discoideum that interact with living yeast particles. Bars in the movies indicate 10 μm.

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Additional file 2:

Movie 2 - Different modes of interaction with budded yeast. The cell expresses PHcrac-GFP (green), a label for phosphoinositides PI(3,4,5)P3 and PI(3,4)P2 and mRFP-LimEΔ (red) for actin filaments. The fluorescence images are superimposed on greyscale brightfield images. This sequence, corresponding to Figure 2a shows cup extension, retraction to the neck, and severing of the particle. Frame to frame interval is 4 seconds. All Additional files are movies showing phagocytes of Dictyostelium discoideum that interact with living yeast particles. Bars in the movies indicate 10 μm.

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Additional file 3:

Movie 3 - Reversible switching between cup extension and retraction. The labels are the same as in Movie 2. This sequence corresponds to Figure 2b. Frame to frame interval is 4 seconds. All Additional files are movies showing phagocytes of Dictyostelium discoideum that interact with living yeast particles. Bars in the movies indicate 10 μm.

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Additional file 4:

Movie 4 - Coronin dynamics relative to actin polymerization and depolymerization. The cell taking up a budded yeast particle expresses GFP-coronin (green) and mRFP-LimEΔ (red). The movie is shown twice: first with the fluorescent labels superimposed on the greyscale brightfield images, subsequently without the brightfield images. During cup extension, coronin localizes to the boundary of the actin layer bordering the cytoplasm. After the completion of uptake, coronin persists during the period of actin disassembly. Images from this sequence are presented in Figure 3a. Frame to frame interval is 2 seconds. All Additional files are movies showing phagocytes of Dictyostelium discoideum that interact with living yeast particles. Bars in the movies indicate 10 μm.

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Additional file 5:

Movie 5 - FRAP of GFP-actin at the neck region of budded yeast and during the extension of a phagocytic cup. The sequence corresponds to Figure 4(d, e). Brightfield illumination is interspersed between the fluorescence images to visualize the position of the particle. Frame to frame interval is 1 second. All Additional files are movies showing phagocytes of Dictyostelium discoideum that interact with living yeast particles. Bars in the movies indicate 10 μm.

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Additional file 6:

Movie 6 - Actin accumulation at the neck region of budded yeast taken up by mutant cells that are deficient in both cortexillin I and II. Complete uptake is shown in the left cell and attempted uptake in the right cell. Frame to frame interval is 1 second. All Additional files are movies showing phagocytes of Dictyostelium discoideum that interact with living yeast particles. Bars in the movies indicate 10 μm.

Format: MOV Size: 174KB Download file

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Additional file 7:

Movie 7 - Accumulation of IBARa-GFP at the neck region of a budded particle. This sequence corresponds to Figure 8d. It shows progression of a phagocytic cup over the neck region and final release of the particle. I-BARa is concentrated for a period of 42 second at the neck, before it disappears there prior to release of the particle. Frame to frame interval is 0.5 seconds. All Additional files are movies showing phagocytes of Dictyostelium discoideum that interact with living yeast particles. Bars in the movies indicate 10 μm.

Format: MOV Size: 2.2MB Download file

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Additional file 8:

Movie 8 - Localization of GFP-MyoB (green) and mRFP-ArpC1 (red) to the phagosome membrane surrounding the neck of a particle. Intense accumulation of these markers persists for 3 minutes, and release of the particle is preceded by their disappearance. Frame to frame interval is 1 second. All Additional files are movies showing phagocytes of Dictyostelium discoideum that interact with living yeast particles. Bars in the movies indicate 10 μm.

Format: MOV Size: 80KB Download file

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Additional file 9:

Movie 9 - Release of a yeast carcass at the end of the phagocytic pathway. This sequence shows the involvement of GFP-MyoB (green) and mRFP-ArpC1 (red) during exocytosis. Frame to frame interval is 1 second. All Additional files are movies showing phagocytes of Dictyostelium discoideum that interact with living yeast particles. Bars in the movies indicate 10 μm.

Format: MOV Size: 544KB Download file

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Additional file 10:

Movie 10 - Recruitment of GFP-myosin-II to a retracting cup. The cell expresses GFP-myosin-II heavy chain (green) and mRFP-LimEΔ (red). Myosin-II does not colocalize with actin at the neck of the particle, but localizes to the lateral border of the cup during its retraction, and to other retracting regions of the cell body. Frame to frame interval is 1 second. All Additional files are movies showing phagocytes of Dictyostelium discoideum that interact with living yeast particles. Bars in the movies indicate 10 μm.

Format: MOV Size: 128KB Download file

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Additional file 11:

Movie 11 - A myosin-II-null cell that severs a budded yeast particle. The cell expressing GFP-LimEΔ is also shown in Figure 11b. Brightfield images are interspersed between the fluorescence recordings to show the particle before and after severing. Frame to frame interval is 1 second. All Additional files are movies showing phagocytes of Dictyostelium discoideum that interact with living yeast particles. Bars in the movies indicate 10 μm.

Format: MOV Size: 902KB Download file

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Open Data