Open AccessProtein Transfection of Intact Microvessels Specifically Modulates Vasoreactivity and Permeability
Author(s) -
John H. Tinsley,
David C. Zawieja,
Mack H. Wu,
Elena E. Ustinova,
Wei Xu,
Sarah Y. Yuan
Publication year - 2001
Publication title -
journal of vascular research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.58
H-Index - 74
eISSN - 1423-0135
pISSN - 1018-1172
DOI - 10.1159/000051077
Subject(s) - protein kinase c , transfection , vasodilation , microbiology and biotechnology , nitric oxide , protein kinase a , vascular permeability , nitric oxide synthase , downregulation and upregulation , agonist , microcirculation , chemistry , signal transduction , biology , kinase , biochemistry , medicine , endocrinology , receptor , gene
Precise regulation of microvascular tone and barrier function is essential for proper coronary perfusion and performance. Agonist-induced alterations in either or both of these functions ultimately lead to microcirculatory dysfunction and cardiac insufficiency. Two important pathways involved in regulating vasomotor response and barrier function are the activation of nitric oxide synthase (NOS) and upregulation of protein kinase C (PKC). To date, studies of these two signaling proteins have relied mainly on pharmacological approaches. Unfortunately, the specificity of various inhibitors can be cause for concern. In order to address this problem, a protein transfection technique we developed for cultured endothelial cells has been modified and applied to isolated, intact coronary microvessels. Our results from green fluorescent protein transfection in arterioles and venules showed that this procedure could be used to introduce proteins into the microvascular wall. By transfecting inhibitor peptides against NOS and PKC into coronary arterioles and venules, we have been able to determine the specific roles of these two enzymes in vasodilation and hyperpermeability responses.