This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Fission yeast (Schizosaccharomyces pombe) is an excellent model for studying cell division and cell morphogenesis. Normal polar growth of these cylindrical cells requires microtubules and polarity factors such as tea1, tea2, tip1 and mod5. Interphase microtubules are arranged in 4-5 cytoplasmic bundles oriented along the cell's axis. The individual microtubules within these bundles are believed to grow and shrink independently, based on fluorescence microscopy data. We are working to show the exact range of microtubule numbers in a bundle, in particular the fine organization both at the cell tips and in the nuclear area where the microtubules seem to originate. For this kind of study electron tomography is the method of choice, since one can reconstruct and investigate a volume with an optimal resolution. We have developed a prescreening technique that enables us to acquire tomograms rapidly but with low resolution. The procedure reveals otherwise invisible microtubules in the semi-thin sections. Thus, we can concentrate our efforts on selected cells where we know that microtubules are present. From fluorescence studies it has been suggested that microtubule bundles overlap with their minus ends in the nuclear region. We are examining the exact polarity of the microtubules using the hook method introduced by Heidemann and McIntosh (1980). In combination with the tomograms acquired in collaboration with the Boulder 3D Lab, we can therefore determine polarity, structure and organization of the individual microtubules, as well as see details, such as whether a microtubule end is capped, blunt or flared end and how this correlates with specific subcellular regions.
| Giddings Jr, Thomas H; Morphew, Mary K; McIntosh, J Richard (2017) Preparing Fission Yeast for Electron Microscopy. Cold Spring Harb Protoc 2017: |
| Zhao, Xiaowei; Schwartz, Cindi L; Pierson, Jason et al. (2017) Three-Dimensional Structure of the Ultraoligotrophic Marine Bacterium ""Candidatus Pelagibacter ubique"". Appl Environ Microbiol 83: |
| Brown, Joanna R; Schwartz, Cindi L; Heumann, John M et al. (2016) A detailed look at the cytoskeletal architecture of the Giardia lamblia ventral disc. J Struct Biol 194:38-48 |
| Saheki, Yasunori; Bian, Xin; Schauder, Curtis M et al. (2016) Control of plasma membrane lipid homeostasis by the extended synaptotagmins. Nat Cell Biol 18:504-15 |
| Höög, Johanna L; Lacomble, Sylvain; Bouchet-Marquis, Cedric et al. (2016) 3D Architecture of the Trypanosoma brucei Flagella Connector, a Mobile Transmembrane Junction. PLoS Negl Trop Dis 10:e0004312 |
| Park, J Genevieve; Palmer, Amy E (2015) Properties and use of genetically encoded FRET sensors for cytosolic and organellar Ca2+ measurements. Cold Spring Harb Protoc 2015:pdb.top066043 |
| McCoy, Kelsey M; Tubman, Emily S; Claas, Allison et al. (2015) Physical limits on kinesin-5-mediated chromosome congression in the smallest mitotic spindles. Mol Biol Cell 26:3999-4014 |
| Höög, Johanna L; Lötvall, Jan (2015) Diversity of extracellular vesicles in human ejaculates revealed by cryo-electron microscopy. J Extracell Vesicles 4:28680 |
| Marc, Robert E; Anderson, James R; Jones, Bryan W et al. (2014) The AII amacrine cell connectome: a dense network hub. Front Neural Circuits 8:104 |
| Weber, Britta; Tranfield, Erin M; Höög, Johanna L et al. (2014) Automated stitching of microtubule centerlines across serial electron tomograms. PLoS One 9:e113222 |
Showing the most recent 10 out of 84 publications
