Endocardial TRPC-6 Channels Act as Atrial Mechanosensors and Load-Dependent Modulators of Endocardial/Myocardial Cross-Talk
Vesna Nikolova-Krstevski, Soeren Wagner, Ze Yan Yu, Charles D. Cox, Jasmina Cvetkovska, Adam P. Hill, Inken G. Huttner, Victoria Benson, Andreas A. Werdich, Calum MacRae, Michael P. Feneley, Oliver Friedrich, Boris Martinac and Diane Fatkin
Effects of Stretch on TRPC-6 Expression and Localization in AE Cells
(A) Fluorescence immunostaining of TRPC-6 (green), F-actin (red), and nuclear 4′,6-diamidino-2-phenylindole (DAPI) stain (blue) in control (nonstretched) atrial endocardium (AE) cells (left column), and in cells stretched for 15 min (middle column) and 24 h (right column). In the control cells and cells subjected to 15-min stretch, TRPC-6 is localized at the cell membrane and other intracellular locations. In cells stretched for 24 h, TRPC-6 accumulates in the cell interior, mostly in and around the nucleus. Scale bar = 20 μm. (B) Western blot evaluation of TRPC-6 protein levels in AE cells stretched for 0, 1, 4, and 24 h. Representative blots (left panel) and quantification from 3 different TRPC-6 immunoblots (right panel) are shown. β-tubulin immunolabeling was used as loading control. See Supplemental Figure 7B for full Western blots. All values are mean ± SEM. *p < 0.001 versus control cells. Abbreviation as in Figure 1.