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Cardiovascular Research 2002 53(2):431-438; doi:10.1016/S0008-6363(01)00496-5
© 2002 by European Society of Cardiology
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Copyright © 2001, European Society of Cardiology

Role of protein kinase C- or RhoA-induced Ca2+ sensitization in stretch-induced myogenic tone

Dong-Soo Yeona, Jung-Sup Kima, Duck-Sun Ahna, Seong-Chun Kwona,b, Bok-Soon Kangc, Kathleen G. Morgand,e and Young-Ho Leea,b,*

aDepartment of Physiology, College of Medicine, Yonsei University, C.P.O. Box 8044, Seoul, 120-752, Republic of Korea
bBK 21 Project for Medical Sciences, Yonsei University, C.P.O. Box 8044, Seoul, 120-752, Republic of Korea
cDepartment of Physiology, College of Medicine, Pochon CHA University, Pochon, 487-800, Republic of Korea
dSignal Transduction Group, Boston Biomedical Research Institute, Watertown, MA 02472, USA
eCardiovascular Division, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, MA 02215, USA

* Corresponding author. Tel.: +82-2-361-5197; fax: +82-2-393-0203 yhlee{at}yumc.yonsei.ac.kr

Objective: It has been suggested that Ca2+ sensitization mechanisms might contribute to myogenic tone. However, specific mechanisms have yet to be fully identified. Therefore, we investigated the role of protein kinase C (PKC)- or RhoA-induced Ca2+ sensitization in myogenic tone of the rabbit basilar vessel. Methods: Myogenic tone was developed by stretch of rabbit basilar artery. Fura-2 Ca2+ signals, contractile responses, PKC immunoblots, translocation of PKC and RhoA, and phosphorylation of myosin light chains were measured. Results: Stretch of the resting vessel evoked a myogenic contraction and an increase in the intracellular Ca2+ concentration ([Ca2+]i) only in the presence of extracellular Ca2+. Stretch evoked greater contraction than high K+ at a given [Ca2+]i. The stretch-induced increase in [Ca2+]i and contractile force were inhibited by treatment of the tissue with nifedipine, a blocker of voltage-dependent Ca2+ channel, but not with gadolinium, a blocker of stretch-activated cation channels. The PKC inhibitors, H-7 and calphostin C, and a RhoA-activated protein kinase (ROK) inhibitor, Y-27632, inhibited the stretch-induced myogenic tone without changing [Ca2+]i. Immunoblotting using isoform-specific antibodies showed the presence of PKC{alpha} and PKC{varepsilon} in the rabbit basilar artery. PKC{alpha}, but not PKC{varepsilon}, and RhoA were translocated from the cytosol to the cell membrane by stretch. Phosphorylation of the myosin light chains was increased by stretch and the increased phosphorylation was blocked by treatment of the tissue with H-7 and Y-27632, respectively. Conclusions: Our results are consistent with important roles for PKC and RhoA in the generation of myogenic tone. Furthermore, enhanced phosphorylation of the myosin light chains by activation of PKC{alpha} and/or RhoA may be key mechanisms for the Ca2+ sensitization associated myogenic tone in basilar vessels.

KEYWORDS Microcirculation; Arteries; Calcium (cellular); E–c coupling; Stretch/m–e coupling


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