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Journal of Histochemistry and Cytochemistry, Vol. 49, 323-332, March 2001, Copyright © 2001, The Histochemical Society, Inc.

ARTICLE

Selective Expression of the Proprotein Convertases Furin, PC5, and PC7 in Proliferating Vascular Smooth Muscle Cells of the Rat Aorta In Vitro

Philipp Stawowya,b, Jadwiga Marcinkiewicza, Kristof Grafb, Nabil G. Seidaha, Michel Chrétienc, Eckart Fleckb, and Mieczyslaw Marcinkiewicza
a Laboratory of Biochemical Neuroendocrinology, Clinical Research Institute of Montreal, Montreal, Quebec, Canada
b Department of Medicine/Cardiology, Charité, Campus Virchow-Clinic, Humboldt-University Berlin and German Heart Institute, Berlin, Germany
c Department of Molecular Medicine and The Disease of Aging, Loeb Health Research Institute, Ottawa, Ontario, Canada

Correspondence to: Mieczyslaw Marcinkiewicz, Lab. of Biochemical Neuroendocrinology, Clinical Research Inst. of Montreal, 110 Pine Avenue West, Montreal, QC, Canada H2W 1R7. E-mail: Marcinm@ircm.qc.ca

The aim of this study was to investigate whether transformation of quiescent vascular smooth muscle cells (VSMCs) into proliferating secretory cells is accompanied by an expression of processing enzymes that activate de novo-synthesized growth factors. Three enzymes belonging to the family of the kexin/subtilisin-like mammalian proprotein convertases (PCs), furin, PC5, and PC7, were found to be upregulated after balloon denudation in vivo. To determine their importance in these cell processes, we investigated their gene regulation using a short-term organ culture system. After incubation of rat aorta for 4 and 24 hr in serum-free medium, we demonstrated a significant induction of VSMC proliferation. The affected subset of VSMCs, positive for {alpha}-smooth muscle actin, also expressed proliferating cell nuclear antigen (PCNA). Our results revealed a parallel upregulation of furin, PC5, and PC7 in PCNA-immunolabeled cells. As a substrate model for comparison with PCs we used nerve growth factor (NGF). NGF is known to be activated by PCs. As shown by Northern blotting analysis, NGF mRNA concentration was significantly increased in cultured explants. NGF was released into the culture medium. In conclusion, both PCs and NGF are coordinately modulated on induction of VSMC proliferation.

(J Histochem Cytohem 49:323–331, 2001)

Key Words: protein processing, aorta organ culture, proliferating cell nuclear antigen, nerve growth factor,, {alpha}-smooth muscle cell actin, immunocytochemistry, co-localization


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