Three-Dimensional Analysis and Computer Modeling of the Capillary Endothelial Vesicular System With Electron Tomography
Document Type
Article
Publication Date
8-1-2012
Description
Objective: We examined the three-dimensional organization of the endothelial vesicular system with TEM tomography of semi-thick sections. Materials and methods: Mouse abdominal muscle capillaries were perfused with terbium to label vesicular compartments open to the luminal surface. The tissue was prepared for TEM and semi-thick (250nm) sections were cut. Dual axis tilt series, collected from +60° to -60° at 1° increments, were acquired in regions of labeled abluminal caveolae. These tomograms were reconstructed and analyzed to reveal three-dimensional vesicular associations not evident in thin sections. Results: Reconstructed tomograms revealed free vesicles, both labeled and unlabeled, in the endothelial cytoplasm as well as transendothelial channels that spanned the luminal and abluminal membranes. A large membranous compartment connecting the luminal and abluminal surfaces was also present. Computer modeling of tomographic data and video animations provided three-dimensional perspectives to these structures. Conclusions: Uncertainties associated with other three-dimensional methods to study the capillary wall are remedied by tomographic analysis of semi-thick sections. Transendothelial channels of fused vesicles and free cytoplasmic vesicles give credence to their role as large pores in the transport of solutes across the walls of continuous capillaries.
Citation Information
Wagner, Roger; Modla, Shannon; Hossler, Fred; and Czymmek, Kirk. 2012. Three-Dimensional Analysis and Computer Modeling of the Capillary Endothelial Vesicular System With Electron Tomography. Microcirculation. Vol.19(6). 477-484. https://doi.org/10.1111/j.1549-8719.2012.00181.x PMID: 22435898 ISSN: 1073-9688