Fig. 8.
Ethyl-1-(4-(2,3,3-trichloroacrylamide)phenyl)-5-(trifluoromethyl)-1H-pyrazole-4-carboxylate reduces intracellular Ca2+ concentration in RYR1-p.R163C/dominant-negative transient receptor potential canonical 6 (TRPC6) skeletal muscle cells. Intracellular Ca2+ concentration was measured in both the right and left vastus lateralis muscle of wild-type/dominant-negative TRPC6 (A) and RYR1-p.R163C/dominant-negative TRPC6 muscles (B). In both, the right leg was superfused with the TRPC3 blocker ethyl-1-(4-(2,3,3-trichloroacrylamide)phenyl)-5-(trifluoromethyl)-1H-pyrazole-4-carboxylate, and Ca2+ was measured a second time, after which the animal was exposed to 2% halothane, and intracellular Ca2+ concentration was measured in both the right and left vastus lateralis muscles a final time. Five mice per genotype, wild-type dominant-negative TRPC6 (left leg), 20 cells; wild-type dominant-negative TRPC6 (left leg)–halothane, 18 cells; wild-type dominant-negative TRPC6 (right leg), 17 cells; wild-type dominant-negative TRPC6 (right leg)–ethyl-1-(4-(2,3,3-trichloroacrylamide)phenyl)-5-(trifluoromethyl)-1H-pyrazole-4-carboxylate, 20 cells; wild-type dominant-negative TRPC6 (right leg)–ethyl-1-(4-(2,3,3-trichloroacrylamide)phenyl)-5-(trifluoromethyl)-1H-pyrazole-4-carboxylate and halothane, 19 cells. RYR1-p.R163C/dominant-negative TRPC6 (left leg), n = 22; RYR1-p.R163C/dominant-negative TRPC6 (left leg)–halothane, 22 cells; RYR1-p.R163C/dominant-negative TRPC6 (right leg), 21 cells; RYR1-p.R163C/dominant-negative TRPC6 (right leg)–ethyl-1-(4-(2,3,3-trichloroacrylamide)phenyl)-5-(trifluoromethyl)-1H-pyrazole-4-carboxylate and halothane, 21 cells; RYR1-p.R163C/dominant-negative TRPC6 (right leg)–ethyl-1-(4-(2,3,3-trichloroacrylamide)phenyl)-5-(trifluoromethyl)-1H-pyrazole-4-carboxylate and halothane, 22 cells. The experimental conditions used are indicated on the horizontal axes. The values are expressed as means ± SD for each condition. Statistical analysis was done using a one-way ANOVA with Tukey’s posttest. *P ≤ 0.05.