Fig. 5

Co-activation of TRPM3 mechanosensitive receptors, Piezo1 mechanosensitive receptors, and TRPV1 channels. (A) Presentation of spike clusters within one experiment. Spikes are plotted by separating negative vs. positive amplitude in the control condition (top panels) and after 50 μM PregS application (bottom panels) in females (left) and males (right). Each individual dot indicates a single spike. Dots with similar colours represent one cluster (i.e. representing one fibre or one group of spikes, as separated by the KlustaKwik method. (B, C) Examples of clusters within one nerve of female (B) and male (C) meninges that were sequentially activated by all three agonists (Ba, Ca), both 5 μM Yoda1 and 50 μM PregS (Bb, Cb) or both 50 μM PregS and 1 μM capsaicin (Bc, Cc), or only 50 μM PregS (Bd, Cd). Spike shapes are depicted for each cluster. (D, E) Pie diagrams demonstrate the averaged percentage of clusters with various neurochemical profiles in females (D) and males (E). Data are presented from 7 and 5 experiments in females and males, respectively. Notice, the difference in co-appearance of responses to 5 μM Yoda1 and 50 μM PregS in females and males. In females, up to 44% clusters were activated by PregS and Yoda1 (23%) or by PregS, Yoda1 and capsaicin (21%), whereas in males, the number of such ‘supermechanosensitive’ fibres was only 24% (PregS and Yoda1 (7%); PregS, Yoda1 and capsaicin (17%)). In males, 7% of clusters showed no response to any agonist, whereas this was the case for only 1% of clusters in females where, in addition, no fibres responded only to 5 μM Yoda1