Piezo-Electrically Driven Mechanical Stimulation of Sensory Neurons

Abstract

Mechanotransduction, the conversion of a mechanical stimulus into a biological response, constitutes the basis of a variety of physiological functions such as the senses of touch, balance, proprioception, blood pressure, and hearing. In vertebrates, mechanosensation is mediated by mechanosensory neurons, whose cell bodies are located in trigeminal and dorsal root ganglia. Here, we describe an in vitro model of mechanotransduction that provides an opportunity to explore the properties of mechanosensitive channels in mammalian sensory neurons. The mechano-clamp method allows applying local force on plasma membrane of whole-cell patch-clamped sensory neurons. This technique uses a mechanical probe driven by a computer-assisted piezoelectric microstage to repeatedly stimulate sensory neurons with accurate control of stimulus strength, duration, and speed.

Piezo2 integrates mechanical and thermal cues in vertebrate mechanoreceptors

“…DRG neurons from 6-to 8-wk-old mice and TG neurons from embryonic day 22 to 24 duck embryos were acutely dissociated as previously described (34,35). Electrophysiological recordings of MA currents from neurons and cell lines were performed at different temperatures in the whole-cell mode in response to indentation with a glass probe (35,67,69). For cell-attached experiments, mechanical stimuli were applied by negative pressure pulses using a high-speed pressure clamp system (70).…”

Transduction of Repetitive Mechanical Stimuli by Piezo1 and Piezo2 Ion Channels

“…Mechanical stimulation was performed as previously described by using a high-speed pressure clamp system (HSPC-1; ALA Scientific Instruments, Farmingdale, NY) to apply negative pressure through the patch pipette or by indenting the cell with a fire-polished glass pipette controlled by a piezo-electric driver (E625 LVPZT Controller/Amplifier; Physik Instrumente) (Coste et al, 2010; Hao et al, 2013b; Lewis and Grandl, 2015). Details of the mechanical stimuli are available in Supplemental Experimental Procedures.…”

Touch, Tension, and Transduction – The Function and Regulation of Piezo Ion Channels

“…In contrast to “stretch”, however, “poke” recruits varying numbers of channels with each indentation depth, leading to inconsistent stimulus-response relationships and often resulting in a lack of response saturation prior to patch rupture [46]. While this method requires relatively large deflections from above to elicit currents from the soma (~5 μm) or neurites (~500 nm), small deflections (~10 nm) of micropillar arrays supporting cells from below can also activate Piezo-mediated currents with high sensitivity [47].…”