Relationship between endothelial cell turnover and permeability to horseradish peroxidase.
The quantitative relations between cell turnover (cell mitosis and death) and macromolecular leakage were studied at the level of individual endothelial cells (ECs) in the thoracic aortae of 32 adult male Sprague-Dawley rats. The experiments were performed on en face preparations of aortic specimens obtained 1, 3, 5 or 10 min after the intravenous administration of horseradish peroxidase (HRP). Mitotic ECs were identified by hematoxylin nuclear staining; dying or dead ECs containing cytoplasmic immunoglobulin G were detected by indirect immunocytochemistry and endothelial leakages to HRP were visualized by light microscopy. The number and size of HRP spots increased with time and the spots fused to form large brown areas in 10 min. Quantitative data on the contributions of EC mitosis and EC death to the transendothelial leakages of HRP were obtained in the same animals. Although mitotic ECs (0.01%) and dying ECs (0.1%) were infrequent in occurrence, the great majority (over 90%) of these ECs were associated with focal HRP uptake. These mitotic and dying ECs, however, accounted for only 17% of the total leakage sites indicating that significant leakage of the 4-5 nm HRP also occurs in normal ECs not morphologically identified as being in mitosis or death. The percentages of leaky spots attributable to mitosis or cell death were greater for the 6 nm albumin and the 22 nm low density lipoprotein (LDL) which probably cannot traverse the normal junctions and use the leaky junctions during cell turnover as the major pathway.
Chen
Y L
YL
Institute of Anatomy, School of Life Science, National Yang-Ming University, Taipei, Taiwan.
Jan
K M
KM
Lin
H S
HS
Chien
S
S
D000818:Animals; D001013:Aorta, Thoracic / Q000166:cytology / Q000378:metabolism; D017209:Apoptosis; D002463:Cell Membrane Permeability; D004730:Endothelium, Vascular / Q000166:cytology* / Q000378:metabolism; D006735:Horseradish Peroxidase / Q000378:metabolism*; D007124:Immunoenzyme Techniques; D008297:Male; D008938:Mitosis; D051381:Rats; D017207:Rats, Sprague-Dawley
D006735:Horseradish Peroxidase