Timelapse data of microtubule behaviours in plant cotyledon cells
Description
Microtubule movies showing microtubule dynamics including the discovery of crossover and severing as shown as supplemental data in the following paper: Wightman, R. and Turner, S.R. (2007), Severing at sites of microtubule crossover contributes to microtubule alignment in cortical arrays. The Plant Journal, 52: 742-751. https://doi.org/10.1111/j.1365-313X.2007.03271.x All the video files are of YFP-labelled microtubules in Arabidopsis thaliana cotyledons. The original movies no longer have native support for playback on modern operating systems. They have been converted to H.264 MPEG-4 and deposited here. There are 6 movies in total: Movie 1. Dynamics of a net-like array. The video clip is taken from a pavement cell (YFP-MBD). Bar = 2 μm. Movie 2. Microtubule (MT) bundles resist lateral movement. An MT leading end (left, red arrow) is forced against an obstructing MT running along the edge of the cell. Initial bending only occurs up to an apparent crossover (right, red arrow). Further bending results in an S–shaped configuration but no bending occurs further back from the crosslink where the MT is part of a bundle (green arrow). Bar = 2 μm. Movie 3. Microtubule (MT) collision results in a lower polymerization rate. This movie is taken from a pavement cell. An MT is identified by a red arrow and is seen to pause whilst it negotiates an obstruction. Bar = 2 μm. Movie 4. Microtubule (MT) severing during MT alignment into an ordered array. MTs were observed in a petiole cell undergoing alignment. MTs growing in an arc are severed at sites marked by white arrows. The posterior portions of two MTs undergo a catastrophe (green arrows) leaving the posterior portions intact (yellow arrows). The upper anterior MT then undergoes a catastrophe whilst the lower MT appears to migrate by hybrid treadmilling. Bar = 2 μm. The Fire look-up table of ImageJ was used to aid visualization of the MTs. Movie 5. Microtubule (MT) severing resulting in a new leading end. A bundled leading end (green arrow) undergoes a severing event further back along the MT (white arrow). The posterior portion of the severed MT then undergoes a catastrophe followed by rescue. Growth at the leading end then occurs along with a change in trajectory. Bar = 5 μm. The Fire look-up table of ImageJ was used to aid visualization of the MTs. Movie 6. Nucleation-associated severing. Two clips where, in each case, a microtubule (MT) is seen to emerge from an existing MT (arrowheads). After a period of growth the lagging end retreats from its site of nucleation. Bar = 2 μm.
Files
Steps to reproduce
Data were all acquired on a Leica DMR fluorescent manual microscope equipped with a SPOT Xplorer camera and HCX PL APO ×100 oil NA 1.4–0.7 objective lens. Cotyledons were dissected and mounted in water on microscope slides with a coverslip. Further details of methodology, including construction of the YFP microtubule reporter, can be found here in the accompanying paper.
Institutions
- University of Cambridge