Differential intracellular trafficking of von Willebrand factor (vWF) and vWF propeptide in porcine endothelial cells lacking Weibel-Palade bodies and in human endothelial cells.
Von Willebrand factor (vWF) is an adhesive protein involved in primary haemostasis virtually absent in the thoracic aorta of swine, an animal model widely used in thrombosis and atherosclerosis. By RT-PCR analysis we show that porcine aortic endothelial cells (PAEC) express the vWF gene, although vWF mRNA levels were 8+/-0.8-fold (p<0.05) or 290+/-8.9-fold (p<0.0001) lower than those in porcine pulmonary artery EC (PPEC) or human aortic EC (HAEC), respectively. Although vWF was rare in the thoracic aorta of swine, vWF propeptide (vWFpp) was present in the endothelium of this artery and in both primary and passaged PAEC. In addition, vWFpp but not vWF was detected in PAEC by Western blot. In PAEC neither vWFpp nor P-selectin immunostaining depicted Weibel-Palade bodies (WPB)-like structures, and acute stimuli (alpha-thrombin or the calcium ionophore A23187) did not increase vWF secretion. vWFpp co-localized with a Golgi marker, that cycles between the stacked Golgi (SG fraction) and earlier compartments of the secretory pathway. Our results confirm that PAEC express very low levels of vWF mRNA and indicate that in these cells, that do not have WPB, vWF and vWFpp have divergent intracellular trafficking pathways.
Royo
Teresita
T
Instituto de Investigación Cardiovascular de Barcelona, CSIC-ICCC-Hospital de la Santa Creu i Sant Pau, Avda. Sant Antoni Maria Claret # 167, Spain.
Martínez-González
José
J
Vilahur
Gemma
G
Badimon
Lina
L
D000818:Animals; D001013:Aorta, Thoracic; D001808:Blood Vessels / Q000648:ultrastructure*; D015152:Blotting, Northern; D015153:Blotting, Western; D002465:Cell Movement; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism* / Q000648:ultrastructure*; D006801:Humans; D007150:Immunohistochemistry; D023421:Models, Animal; D011498:Protein Precursors / Q000032:analysis / Q000378:metabolism*; D011651:Pulmonary Artery; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D013045:Species Specificity; D013552:Swine; D020302:Weibel-Palade Bodies / Q000378:metabolism*; D014841:von Willebrand Factor / Q000032:analysis / Q000378:metabolism*
D011498:Protein Precursors; D014841:von Willebrand Factor; C067003:von Willebrand factor propolypeptide