Fig. 2: EC-specific Piezo1 gain-of-function (GOF) mice demonstrate impaired functional hyperemia.

a Crossbreeding scheme used to generate inducible, EC-specific Piezo1-GOF mice. Tamoxifen-treated Cre+ mice are used as GOF, and littermate Cre- mice are the control. b Representative traces of Piezo1 currents recorded from freshly isolated ECs from the somatosensory cortices of control and GOF mice. The cell-attached configuration was used, where EC patches were held at -50 mV in the absence or presence of Yoda1 (5 µM) in the pipette solution. C denotes closed channel. c Averaged data of the open probability (NP_O) of Piezo1 in the absence (n = 11 ECs/6 control; 10 ECs/4 GOF mice) or presence of Yoda1 (n = 10 ECs/6 control; 7 ECs/4 GOF mice). d Schematic of the LSCI setup for imaging blood flow through a thinned skull. (e) The Allen mouse brain atlas highlighting the somatosensory barrel cortex. f, g Differential maps of blood flow responses in a control (f) and a GOF (g) mouse. Whisker stimulation using air puffs (5 Hz, 30 s) evoked hyperemia in the contralateral somatosensory cortex (arrows). h Representative traces and summary data of the hyperemic response to 30 s-whisker stimulations in control (n = 8) and GOF (n = 9) mice. Right: area under the curve analyses (n = 19 whisker stimulations/6 control; 20 stimulations/8 GOF mice). i Representative responses and scatter plots of FH evoked by 5 s air puffs (n = 7 control, n = 9 GOF mice). Area under the curve analysis was calculated from 41 and 43 stimulations from 7 control and 9 GOF mice, respectively. j Representative images demonstrating the staining of ECs (caveolin-1), basement membranes (collagen IV), and nuclei (DAPI) in brain slices of the cortex of GOF and control mice. Scale bar: 100 µm. Vascular densities and string vessel densities in Piezo1^cx/cx;Cdh5-Cre+ (n  =  12) and control (n = 10) mice. k Representative images and averaged data as in j from brain slices of the hippocampus of Piezo1^cx/cx;Cdh5-Cre+ (n = 12) and control (n =10) mice. All statistical tests were unpaired Student’s t test (two-sided, *P < 0.05, **P < 0.01, ***P < 0.001). All error bars are SEM. Scatter plot data in c, h, I, j, k are presented as mean ± SEM. Area under the curve data in h and i are presented as histograms. Source data are provided as a Source Data file.