Mechanosensitive ion channel Piezo2 is inhibited by D-GsMTx4

Abstract

Enterochromaffin () cells are the primary mechanosensors of the gastrointestinal (GI) epithelium. In response to mechanical stimuli cells release serotonin (5-hydroxytryptamine; 5-HT). The molecular details of cell mechanosensitivity are poorly understood. Recently, our group found that human and mouse cells express the mechanosensitive ion channel Piezo2. The mechanosensitive currents in a human cell model QGP-1 were blocked by the mechanosensitive channel blocker D-GsMTx4. In the present study we aimed to characterize the effects of the mechanosensitive ion channel inhibitor spider peptide D-GsMTx4 on the mechanically stimulated currents from both QGP-1 and human Piezo2 transfected HEK-293 cells. We found co-localization of 5-HT and Piezo2 in QGP-1 cells by immunohistochemistry. QGP-1 mechanosensitive currents had biophysical properties similar to dose-dependently Piezo2 and were inhibited by D-GsMTx4. In response to direct displacement of cell membranes, human Piezo2 transiently expressed in HEK-293 cells produced robust rapidly activating and inactivating inward currents. D-GsMTx4 reversibly and dose-dependently inhibited both the potency and efficacy of Piezo2 currents in response to mechanical force. Our data demonstrate an effective inhibition of Piezo2 mechanosensitive currents by the spider peptide D-GsMTx4.

A population of gut epithelial enterochromaffin cells is mechanosensitive and requires Piezo2 to convert force into serotonin release

“…3 D ). The NeuroD1 + cell mechanosensitive current biophysical properties were consistent with Piezo2 ( 13 , 23 , 24 ). We attempted to make a NeuroD1 + cell-specific Piezo2 knockout but NeuroD1-cre ; Piezo2 f/f mating resulted in a lethal phenotype ( SI Appendix , Fig.…”

“…Therefore, we used pharmacological inhibitors and Piezo2 knockdown to test whether the NeuroD1 + cell mechanosensitive currents are Piezo2. In voltage-clamped primary NeuroD1 + cells stimulated by membrane displacement, we found that mechanosensitive currents were inhibited by Gd 3+ , an established mechanosensitive ion channel blocker ( 23 , 26 ) and D-GsMTx4, a Piezo1 ( 27 ) and Piezo2 ( 24 ) blocker ( Fig. 3 E and F ).…”

“…EC cells, the most common type of EE cells ( 16 ), were proposed to be specialized mechanosensors that in response to force release 5-HT ( 1 , 2 ), which stimulates secretory reflexes ( 3 , 4 ). However, EC cell mechanosensitivity was inferred from studies on intact tissues ( 2 , 35 ) and cell models ( 9 – 11 , 24 ). In this study we addressed critical knowledge gaps: whether EE and EC cells are mechanosensitive and, if so, what is the mechanism of their mechanosensitivity?…”

GsMTx4 reduces the reflex pressor response during dynamic hindlimb skeletal muscle stretch in decerebrate rats

“…) and piezo 2 (Alcaino et al. ) channels but not the potassium selective mechanically activated channel TREK‐1 (Bae et al. ).…”

Chronic femoral artery ligation exaggerates the pressor and sympathetic nerve responses during dynamic skeletal muscle stretch in decerebrate rats

“…Specifically, GsMTx4, a relatively selective antagonist for mechano-gated piezo channels (2,6), reduced the pressor and renal sympathetic nerve responses during dynamic hindlimb muscle contraction to a greater extent in "ligated" versus "freely perfused" rats (12). Femoral artery ligation did not, however, increase piezo1 or piezo2 channel protein expression in L 4 and L 5 dorsal root ganglia (DRG) tissue which suggested that an acute and/or chronic sensitization of mechanoreceptors accounted for the exaggerated mechanicallysensitive component of the exercise pressor reflex rather than increased mechanoreceptor expression.…”