Endoglin co-expression with eNOS, SMAD2 and phosphorylated SMAD2/3 in normocholesterolemic and hypercholesterolemic mice: an immunohistochemical study.
Endoglin, a homodimeric transmembrane glycoprotein, is a part of the transforming growth factor-beta (TGF-beta) receptor cascade. It has been demonstrated that endoglin can affect TGF-beta signaling and eNOS expression by affecting SMAD proteins in vitro. We planned to go one step forward and evaluate whether endoglin is co-expressed with SMAD2, phosphorylated SMAD2/3 protein and eNOS in endothelium of normocholesterolemic C57BL/6J mice, and in advanced atherosclerotic lesions in hypercholesterolemic apoE/LDLr-deficient mice by means of fluorescence immunohistochemistry. Female C57BL/6J mice were fed with a chow diet (standard laboratory diet) for 12 weeks after weaning (at the age of 4 weeks). Two-month-old female apoE/LDLr-deficient mice were fed the western type diet (atherogenic diet) containing 21% fat (11% saturated fat) and 0.15% cholesterol for 2 months. Immunohistochemical analysis of endoglin, SMAD2, phosphorylated SMAD2/3 and eNOS expression was performed in mice aortic sinus. Immunohistochemical analysis showed the expression of endoglin in intact endothelium in both C57BL/6J and apoE/LDLr-deficient mice and in endothelium covering the atherosclerotic lesion in apoE/LDLr-deficient mice. Fluorescence immunohistochemistry revealed co-expression of endoglin with SMAD2, phosphorylated SMAD2/3 and eNOS in intact aortic endothelium in C57BL/6J mice. Moreover, strong co-localization of endoglin, SMAD2, phosphorylated SMAD2/3 and eNOS was also detected in endothelium covering atherosclerotic lesions in apoE/LDLr-deficient mice. In conclusion, we suggest that endoglin, SMAD2, phosphorylated SMAD2/3 and eNOS may be important in vessel endothelium homeostasis underlying their role in atherogenesis.
Department of Biological and Medical Sciences, Faculty of Pharmacy in Hradec Kralove, Charles University in Prague, Heyrovskeho, Hradec Kralove, Czech Republic. petr.nachtigal@faf.cuni.cz
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Nachtigal
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Vecerova
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Pospisilova
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Micuda
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Brcakova
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Navarro Hernandez
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Pospechova
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Semecky
D000818:Animals; D001011:Aorta / Q000166:cytology; D001057:Apolipoproteins E / Q000172:deficiency / Q000235:genetics; D004036:Diet, Atherogenic; D000071063:Endoglin; D004730:Endothelium, Vascular / Q000378:metabolism; D005260:Female; D019085:Fluorescent Antibody Technique, Direct; D006937:Hypercholesterolemia / Q000378:metabolism*; D007150:Immunohistochemistry; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism*; D010766:Phosphorylation; D051899:Smad2 Protein / Q000235:genetics / Q000378:metabolism*; D051900:Smad3 Protein / Q000235:genetics / Q000378:metabolism*
D001057:Apolipoproteins E; D000071063:Endoglin; C502004:Eng protein, mouse; D047908:Intracellular Signaling Peptides and Proteins; D051899:Smad2 Protein; D051900:Smad3 Protein; D052250:Nitric Oxide Synthase Type III