|
Project properties |
|
| Title | Does the formation of cytoplasmic strands depend on actin? |
| Group | Plant Cell Biology, Laboratory of |
| Project type | thesis |
| Credits | 18-39 |
| Supervisor(s) | Hannie van der Honing, Tijs Ketelaar |
| Examiner(s) | Prof. dr. Marcel Janson |
| Contact info | tijs.ketelaar@wur.nl, 0317-482453 |
| Begin date | 2008/04/01 |
| End date | 2010/04/01 |
| Description |
Proper intracellular organization is of vital importance for cellular functioning and plant development. The cytoplasm of plant cells is organized such that a pool of cytoplasm, present around the nucleus and a pool of cytoplasm in the periphery of the cell are connected by strands of cytoplasm. These cytoplasmic strands penetrate the vacuole that separates the two pools of cytoplasm. Cytoplasmic strands are essential for the transport of transcripts, proteins, and signals from the nucleus to the periphery of the cell and vice versa. All cytoplasmic strands of interphase plant cells contain actin filaments, and when actin filaments are depolymerized, the strands disappear. Thus, actin filaments are the structural basis of cytoplasmic strands. it is likely that actin filaments are also involved in the formation of cytoplasmic strands, but has not been proven yet that this is the case. In this MSc project, you will use tobacco BY-2 suspension cells that express GFP, targeted to the vacuolar (or tonoplast) membrane (GFP:VacM) to visualize the boundaries of cytoplasmic strands and BY-2 cells expressing GFP:FABD2, which decorates the actin cytoskeleton. We can experimentally remove cytoplasmic strands by treatment with the actin depolymerising drug latrunculin B. After removal of latrunculin B, cells recover and cytoplasmic strands re-form. You will follow this recovery over time by 4D (time, z) imaging. By analysis of these data, we will get insight in the formation of cytoplasmic strands. We will also perform this experiment in the presence of microtubule depolymerizing drugs, to exclude the possibility of microtubules playing a role in the formation of cytoplasmic strands. By using BY-2 cells in which expression of specific actin interacting proteins is inhibited (RNAi), we can test the role of actin binding proteins in cytoplasmic strand formation. In this project, we will focus on actin bundling proteins. |
| Used skills | Culturing of tobacco BY-2 suspension cells
Cell synchronization Application of cytoskeleton drugs Microscopy techniques (DIC, CLSM and spinning disk confocal microscopy) Digital image analysis Molecular cloning, RNAi |
| Requirements | PCB-30306 |