Adventitial remodeling after angioplasty is associated with expression of tenascin mRNA by adventitial myofibroblasts
Kurt Wallner, MD*,
Behrooz G. Sharifi, PhD*,
Prediman K. Shah, MD, FACC*,
Sumiko Noguchi, MD ,
Hector DeLeon, MD, PhD and
Josiah N. Wilcox, PhD
* Atherosclerosis Research Center, Division of Cardiology, Burns and Allen Research Institute, Cedars-Sinai Medical Center, UCLA School of Medicine, Los Angeles, California,, USA
Department of Medicine, Emory University School of Medicine, Atlanta, Georgia, USA
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Figure 1 Immunohistochemical localization of TnC in rat carotid arteries. Frozen sections of uninjured (A) as well as arteries collected two days (B), three days (C), four days (D), five days (E) and seven days (F) after injury were stained with a polyclonal anti-TnC antibody (1:500). Adjacent sections were stained with control preimmune antibodies. The lumen is toward the top of each panel. All photomicrographs are at a x100 magnification. Substitution of anti-TnC antibodies with an equivalent concentration of preimmune rabbit serum (1:500) or an irrelevant antibody produced no staining of normal or injured carotid artery sections. adv = adventitia; m = media; TnC = tenascin-C.
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Figure 2 In situ expression of TnC in injured rat arteries. TnC mRNA was detected by in situ hybridization 3 (A) or 14 (B) days after balloon injury of rat carotid artery using full-length TnC riboprobes. Cryosections were treated with proteinase K. After hybridization, the sections were washed, dried, coated and exposed in the dark at 4°C for four weeks. After development, the sections were counterstained with hematoxylin and eosin to aid in cell identification and viewed on a photomicroscope. TnC = tenascin-C.
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Figure 3 Localization of tenascin-C by in situ hybridization (A and C) and immunohistochemistry (B and D) after balloon overstretch injury of porcine coronary arteries. Balloon injury was performed in porcine left anterior descending coronary arteries as described in the Methods section and injured vessels harvested 3 (A and B) or 14 days (C and D) after injury. adv = adventitia; m = media.
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Figure 4 Effect of angiotensin II and PDGF-BB on TnC mRNA and protein expression. Quiescent culture of porcine adventitial cells were treated with 1 nM PDGF-BB or 100 µM angiotensin II for the indicated times (in hours). Total RNA was extracted and analyzed by Northern blot and hybridized with TnC. To control for loading, the blot was rehybridized with the GAPDH cDNA probe. (B) For Western blot analysis, the media from cultured cells treated with angiotensin II or PDGF-BB for 8 and 24 h were collected and analyzed using 1:5,000 dilution of chicken antibodies to the recombinant fibrinogen-like domain of TnC. GAPDH = glyceraldehyde-phosphate dehydrogenase; TnC = tenascin-C.
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