title: Vacuole formation in the endothelium of rat extremity vessels depends on fixation techniques and vessel type.
abstract: Applying immersion fixation for electron microscopy, huge clear endothelial membrane-bound vacuoles of 0.1-3 microm diameter were noted in the extremity veins of Sprague-Dawley rats. Histological and electron microscopic histochemical methods were applied to determine whether they were the product of programmed cell death or any other kind of cell damage. Image analyzer was used to measure the total area of the vacuoles in the endothelium cells. Neither lipid content nor acidic phosphatase activity could be identified in the vacuoles. In saphenous and brachial veins, the vacuoles occupied 20.6 +/- 2.21% and 18 +/- 2.45% of the endothelium, respectively. Venous endothelium of two different strains of rat also contained the vacuoles. No such structures appeared in extremity arteries. Long-term tilting did not influence vacuolization. Using in vivo whole body fixation, only pinocytotic and dense microvesicles, but no vacuoles were noted. In conclusion, the clear vacuolar structures represent neither lipid inclusions nor secondary lysosomes. The method of tissue fixation is critical when venous endothelial vesicles are investigated. It is presumed that the vacuoles originated from intra- or intercellular microstructures, and that in case of the collapsible vein segments, their size is increased under the pathological-hypoxic and low-pressure-conditions of in vitro fixation.
authors.affiliation: Institute of Human Physiology and Clinical Experimental Research, Semmelweis University Budapest, Ullõi út 78a, PO Box 448, Budapest 1082, Hungary. raffai@elet2.sote.hu
authors.forename: Gábor
authors.initials: G
authors.lastname: Raffai
authors.forename: Erzsébet
authors.initials: E
authors.lastname: Fehér
authors.forename: György
authors.initials: G
authors.lastname: Nádasy
authors.forename: Sándor
authors.initials: S
authors.lastname: Paku
authors.forename: Gábor
authors.initials: G
authors.lastname: Pogány
authors.forename: Ferenc
authors.initials: F
authors.lastname: Tímár
authors.forename: Béla
authors.initials: B
authors.lastname: Szende
authors.forename: Emil
authors.initials: E
authors.lastname: Monos
mesh_terms: D000818:Animals; D016477:Artifacts; D001916:Brachial Artery / Q000648:ultrastructure; D022162:Cytoplasmic Vesicles / Q000648:ultrastructure*; D042783:Endothelial Cells / Q000648:ultrastructure*; D005121:Extremities / Q000098:blood supply; D008297:Male; D008854:Microscopy, Electron; D051381:Rats; D011916:Rats, Inbred F344; D017207:Rats, Sprague-Dawley; D012501:Saphenous Vein / Q000648:ultrastructure; D016707:Tissue Fixation; D014617:Vacuoles / Q000648:ultrastructure*