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. The Kadler laboratory is focused on understanding how mesenchymal cells deposit an extracellular matrix comprising spatially organized arrays of collagen fibrils. We recently reported on a novel protein secretion pathway, the 'fibripositor pathway'that is utilized by embryonic tendon fibroblasts to deposit parallel bundles of collagen fibrils to intercellular channels. The key features of the pathway are intracellular cleavage of procollagen (the precursor of collagen) to collagen in the late secretory pathway, the assembly of early collagen fibrils in Golgi-to-plasma membrane carriers (GPCs) and export of the fibrils through specialized plasma membrane structures called fibripositors. In February of this year, Drs. David Holmes, Toby Starborg and Karl Kadler visited the 3-D lab to use their electron tomography instrument to determine the structure of single fibripositors and the GPCs that contain early collagen fibrils. During our 6-day visit we collected 26 tilt series to generate tomograms from orthogonal dual axis tilt series. Our ability to collect such excellent data was in large part attributable to the excellent support and scientific input we obtained from Dr. Eileen O'Toole, who operated the microscope and provided instruction on how to use the tomography software. These data sets contain new information on the 3D structure of collagen fibril bundles including interfibrillar filaments. Each of these data sets is likely to lead to separate research articles in 2005 and 2006. These data are unique to our research group. They provide us with an unrivalled opportunity to discover the mechanism of cell-mediated synthesis of the extracellular matrix, and the role of the cytoskeleton in determining cell shape and fibripositor assembly.
| 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 |
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