title: The vesiculo-vacuolar organelle (VVO): a distinct endothelial cell structure that provides a transcellular pathway for macromolecular extravasation.
abstract: The vesiculo-vacuolar organelle (VVO) is a recently described organelle found in the cytoplasm of endothelial cells that line tumor microvessels and normal venules. VVOs are grape-like clusters of interconnecting uncoated vesicles and vacuoles, bounded by trilaminar unit membranes, that span the entire thickness of vascular endothelium, thereby providing a potential trans-endothelial connection between the vascular lumen and the extravascular space. Macromolecular tracers preferentially cross hyperpermeable tumor microvessels through VVOs. The present investigation was undertaken to elucidate further the ultrastructure and function of VVOs in a murine ovarian carcinoma (MOT) and in normal venules. Morphometry revealed that VVOs were enormous cytoplasmic structures (median area, 0.12-0.14 microns2 in single electron micrographs). Moreover, the individual vesicles and vacuoles that comprised VVOs were on average substantially larger than capillary caveolae and followed a non-normal distribution that was skewed to the right. Specimen tilting provided conclusive evidence that individual VVO vesicles and vacuoles communicated with each other and with the endothelial cells' plasma membranes by stomata, some of which were closed by diaphragms composed of a single membrane. Studies with two tracers, ferritin (FE, diameter approximately 11 nm) and horseradish peroxidase (HRP, diameter approximately 5 nm), revealed that passage of macromolecules through VVOs was regulated at the level of stomatal diaphragms, thereby demonstrating a mechanism for controlling the passage of macromolecules across endothelial cells. Thus, compared with tumor microvessels, little circulating FE and HRP entered the VVOs of normal venular endothelium because stomata joining vesicles and vacuoles to each other and to the lumen and ablumen were closed. VVOs and their component vesicles/vacuoles were readily distinguished from endosomal organelles such as coated vesicles and multivesicular bodies, which also accumulated FE and HRP. Our findings indicate that VVOs provide a major pathway for the extravasation of circulating macromolecules across endothelia taller than capillary endothelium and suggest that upregulated VVO function accounts for the well-known hyperpermeability of tumor blood vessels.
authors.affiliation: Department of Pathology, Beth Israel Hospital, Boston, MA 02215, USA.
authors.forename: A M
authors.initials: AM
authors.lastname: Dvorak
authors.forename: S
authors.initials: S
authors.lastname: Kohn
authors.forename: E S
authors.initials: ES
authors.lastname: Morgan
authors.forename: P
authors.initials: P
authors.lastname: Fox
authors.forename: J A
authors.initials: JA
authors.lastname: Nagy
authors.forename: H F
authors.initials: HF
authors.lastname: Dvorak
mesh_terms: D000818:Animals; D002199:Capillary Permeability; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure; D018699:Coated Vesicles / Q000378:metabolism* / Q000648:ultrastructure; D011992:Endosomes / Q000378:metabolism; D004730:Endothelium, Vascular / Q000378:metabolism / Q000648:ultrastructure*; D005119:Extravasation of Diagnostic and Therapeutic Materials / Q000378:metabolism*; D005260:Female; D005293:Ferritins / Q000493:pharmacokinetics*; D006735:Horseradish Peroxidase / Q000493:pharmacokinetics*; D046911:Macromolecular Substances; D051379:Mice; D008809:Mice, Inbred C3H; D008854:Microscopy, Electron; D010051:Ovarian Neoplasms / Q000648:ultrastructure; D014617:Vacuoles / Q000378:metabolism* / Q000648:ultrastructure
chemical_list: D046911:Macromolecular Substances; D005293:Ferritins; D006735:Horseradish Peroxidase