title: Tubular profiles do not form transendothelial channels through the blood-brain barrier.
abstract: The contribution of tubular profiles within the mammalian cerebral endothelium to the formation of transcellular channels was analysed following exposure of the endothelium to native horseradish peroxidase (HRP) dissolved in saline or dimethyl sulphoxide (DMSO) administered intravenously in mice. Within 5-15 min, but not at 30 min to 2 h postinjection, peroxidase-positive extravasations were evident within the parenchyma of the forebrain and brainstem of mice exposed and not exposed to DMSO. The extravasations may be associated with the rupture of interendothelial tight junctions at the level of arterioles as a consequence of the perfusion-fixation process. Ultrastructural inspection of endothelia within and away from areas of peroxidase extravasation revealed the following intraendothelial, peroxidase-positive organelles: presumptive endocytic vesicles, endosomes (a prelysosomal compartment), multivesicular and dense bodies, and tubular profiles. Statistical analysis of the concentration of HRP-labelled presumptive endocytic vesicles, which may coalesce to form tubules, within endothelia from mice injected intravenously with HRP-DMSO compared to mice receiving HRP-saline revealed no significant difference. HRP-positive tubular profiles were blunt-ended, variable in length and width, and appeared free in the cytoplasm or in continuity with dense bodies. Labelled tubules free in the cytoplasm were positioned parallel to the luminal and abluminal plasma membranes and were less frequently oblique or perpendicular to these membranes. Tubular profiles analysed in serial thin sections or with a goniometer tilt stage did not establish membrane continuities with the luminal and abluminal plasma membranes. Peroxidase-positive tubular profiles were similar morphologically to those exhibiting acid hydrolase activity but did not share morphological and enzyme cytochemical similarities with the endoplasmic reticulum that stained for glucose-6-phosphatase (G6Pase) activity. G6Pase-positive profiles of endoplasmic reticulum were not observed to contribute to a transendothelial canalicular network. Our results suggest that: (i) peroxidase-labelled tubules, acid hydrolase-positive tubules, and G6Pase-positive endoplasmic reticulum do not form transcellular channels through the cerebral endothelium; (ii) tubular profiles labelled with blood-borne HRP in the cerebral endothelium are associated with the endosome apparatus and/or the lysosomal system of organelles; and (iii) DMSO does not appear to alter the permeability of the blood-brain barrier to blood-borne protein.
authors.affiliation: Department of Pathology, University of Maryland School of Medicine, Baltimore 21201.
authors.forename: B J
authors.initials: BJ
authors.lastname: Balin
authors.forename: R D
authors.initials: RD
authors.lastname: Broadwell
authors.forename: M
authors.initials: M
authors.lastname: Salcman
mesh_terms: D000818:Animals; D001692:Biological Transport; D001812:Blood-Brain Barrier / Q000187:drug effects*; D001921:Brain / Q000378:metabolism; D002199:Capillary Permeability / Q000187:drug effects*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000648:ultrastructure*; D005260:Female; D006651:Histocytochemistry; D006735:Horseradish Peroxidase / Q000378:metabolism; D008297:Male; D051379:Mice; D008854:Microscopy, Electron; D009940:Organoids / Q000648:ultrastructure
chemical_list: D006735:Horseradish Peroxidase; D004121:Dimethyl Sulfoxide