Use of Cell-Stretch System to Examine the Characteristics of Mechanosensor Channels: Axonal Growth/Neuroregeneration Studies

Temperature-sensitive TRP (so-called “thermoTRP”) channels are well recognized for their contributions to sensory transduction, responding to a wide variety of stimuli including temperature, nociceptive stimuli, touch, and osmolarity. However, the precise roles for the thermoTRP channels during development have not been determined. To explore the functional importance of thermoTRP channels during neural development, the temporal expression was determined in embryonic mice. Interestingly, TRPV2 expression was detected in spinal motor neurons in addition to the DRG from E10.5, and was localized in axon shafts and growth cones, suggesting that the channel is important for axon outgrowth regulation. We revealed that endogenous TRPV2 was activated in a membrane-stretch-dependent manner in developing neurons by knocking down the TRPV2 function with dominant negative TRPV2 and TRPV2-specific shRNA, and significantly promoted axon outgrowth. In this section, the author introduces experimental methods to investigate the mehcanosensor functions of TRPV2 in axonal outgrowth or regeneration.