Premium
Effect of cell passage and density on protein kinase G expression and activation in vascular smooth muscle cells
Author(s) -
Lin Guiting,
Chow Sylvia,
Lin Jackie,
Wang Guifang,
Lue Tom F.,
Lin ChingShwun
Publication year - 2004
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.20043
Subject(s) - blot , phosphoprotein , smooth muscle , cgmp dependent protein kinase , vascular smooth muscle , immunofluorescence , chemistry , microbiology and biotechnology , cell , phosphorylation , protein kinase a , biology , endocrinology , immunology , biochemistry , antibody , cyclin dependent kinase 2 , gene
It has been shown that rat aortic smooth muscle cells (AoSMCs) lost PKG‐I expression when propagated repetitively or grown at low densities. Conversely, AoSMCs isolated from PKG‐I deficient mice are indistinguishable from those isolated from normal mice in morphology and growth characteristics. In this study, human AoSMCs were grown from passage 9 (p9) to passage 15 (p15) and rat AoSMCs were isolated and cultured from p1 through p15. Western blotting and immunofluorescence microscopy showed little difference in PKG‐I expression among different passages. Next, rat AoSMCs of p4 were grown and harvested at different cell densities. Western blotting again showed little difference among cells seeded or harvested at different densities. To test the effect of cell passage on PKG‐I activation, rat AoSMCs of p4 and p11 were treated with cGMP and analyzed by Western blotting for phosphorylated vasodilator‐stimulated phosphoprotein (P‐VASP). The results showed that p4 had higher level of PKG‐I activation than p11. © 2004 Wiley‐Liss, Inc.