Figure 2. Diet-induced obesity impairs AKAP150_EC-PKC-TRPV4_EC signaling and vasodilation.

(A) Resting MAP (mm Hg) averaged over 3 days in HFD and NFD mice (n = 5; **P < 0.01 for HFD vs. NFD; t-test). (B) Representative diameter traces (left) and averaged diameter data (right) for CCh (0.3–10 μM)-induced dilation of MAs from NFD and HFD mice (n = 8–11; ***P < 0.001 for 1 μM, 3 μM, and 10 μM CCh for NFD vs. HFD mice; two-way ANOVA). (C) Representative diameter traces (left) and averaged diameter data (right) for GSK101 (3–30 nM)-induced vasodilation in NFD and HFD mice (n = 5–9; P < 0.01 [3 nM GSK10] and < 0.001 [10 nM GSK101]; two-way ANOVA). (D) Representative F/F₀ traces from TRPV4_EC sparklet sites (20 μM CPA + GSK101, 10 nM) at MEPs in Fluo-4–loaded en face preparations of MAs from NFD (left) and HFD (right) mice. Dotted red lines indicate quantal levels. (E) Averaged TRPV4_EC sparklet activity (NP_O) at MEPs in MAs from NFD and HFD mice under baseline conditions (20 μM CPA) or in response to GSK101 (10 nM) (n = 7; sparklet activity: *P < 0.05 [CPA] and < 0.01 [GSK101] for NFD vs. HFD). (F) Averaged TRPV4_EC sparklet activity in MAs from NFD and HFD mice under baseline conditions (20 μM CPA) and in response to GSK101 (10 nM) (n = 7). (G) Representative diameter traces for CCh (0.3–10 μM)-induced dilation of splenius muscle arteries from non-obese and obese individuals. (H) Representative diameter traces for GSK101 (3–30 nM)-induced dilation of splenius muscle arteries from obese and non-obese individuals. (I) Averaged diameter data for CCh -induced vasodilation in human splenius and temporalis muscle arteries from obese and non-obese individuals (n = 6; P< 0.01 vs. non-obese; two-way ANOVA). (J) Averaged diameter data for GSK101-induced vasodilation in human splenius and temporalis muscle arteries from obese and non-obese individuals (n = 6; P < 0.01, < 0.01, vs. non-obese; two-way ANOVA). (K) Greyscale image of a field of view with ~ 20 ECs from an en face preparation of Fluo-4 loaded splenius muscle artery (SMA) from a non-obese individual; yellow squares indicate sparklet sites in with CPA + GSK101 (10 nM). (L) TRPV4_EC sparklet activity (NP_O) at MEP in SMAs from non-obese and obese individuals in response to GSK101 (10 nM) and in the presence of GSK219 (100 nM) (n = 4; P < 0.001 [GSK101] for non-obese vs. obese and P < 0.01 [GSK101] vs. [GSK219]; one-way ANOVA).