Transcytosis of albumin in capillary endothelium.
We have used a variety of immunocytochemical procedures to localize albumin in transit through the capillary endothelium of the murine myocardium and thereby identify endothelial cell structures involved in albumin efflux. The most informative results were obtained with a protocol that included (a) removal of endogenous albumin by perfusion of the heart with PBS supplemented with 14 mM glucose, (b) perfusion of the heart vasculature with exogenous (bovine) albumin for various short time periods, (c) fixation of the vessels by formaldehyde-glutaraldehyde mixtures, (d) processing of fixed myocardium specimens through L. R. White embedding followed by sectioning, or direct thin frozen sectioning, and (e) reacting the surface of such specimens with antialbumin antibodies followed by gold-labeled secondary antibodies. The results indicate that (a) monomeric albumin binds (with low affinity) to the luminal surface of the capillary endothelium, (b) it is restricted in transit through the endothelium to plasmalemmal vesicles, and (c) it appears in the pericapillary spaces less than 15 s after the beginning of its perfusion. No albumin concentration gradients, centered with their maxima on the exits from intercellular spaces, were detected at any time points, including the shortest ones (15 and 30 s) investigated. Additional information comparing monomeric vs. polymeric albumin transcytosis was obtained using albumin-gold complexes. The results are discussed in terms of vesicular transport of albumin across the endothelium and the relations of this type of transport to the postulated pore systems of the physiological literature.
Milici
A J
AJ
Department of Surgery, Yale University School of Medicine, New Haven, Connecticut 06510.
Watrous
N E
NE
Stukenbrok
H
H
Palade
G E
GE
D000818:Animals; D001692:Biological Transport; D002196:Capillaries / Q000378:metabolism / Q000648:ultrastructure; D002462:Cell Membrane / Q000648:ultrastructure; D003326:Coronary Circulation; D004730:Endothelium, Vascular / Q000378:metabolism* / Q000648:ultrastructure; D007124:Immunoenzyme Techniques; D008297:Male; D051379:Mice; D008854:Microscopy, Electron; D009206:Myocardium / Q000378:metabolism / Q000648:ultrastructure; D012710:Serum Albumin, Bovine / Q000378:metabolism*
D012710:Serum Albumin, Bovine