Localized and non-contact mechanical stimulation of dorsal root ganglion sensory neurons using scanning ion conductance microscopy

Daniel Sánchez, Uma Anand, Julia Gorelik, Christopher D Benham, Chas Bountra, Max Lab, David Klenerman, Rolfe Birch, Praveen Anand, Yuri Korchev

    Research output: Contribution to journalArticlepeer-review

    39 Citations (Scopus)

    Abstract

    Mechanosensitive ion channels convert external mechanical force into electrical and chemical signals in cells, but their physiological function in different tissues is not clearly understood. One reason for this is that there is as yet no satisfactory physiological method to stimulate these channels in living cells. Using the nanopipette-probe of the Scanning Ion Conductance Microscope (SICM), we have developed a new technique to apply local mechanical stimulus to living cells to an area of about 0.385 microm2, determined by the pipette diameter. Our method prevents any physical contact and damage to the cell membrane by use of a pressure jet applied via the nanopipette. The study used whole-cell patch-clamp recordings and measurements of intracellular Ca2+ concentration to validate the application of the mechanical stimulation protocols in human and rat dorsal root ganglia (DRG) sensory neurons. We were able, for the first time, to produce a non-contact, controlled mechanical stimulation on living neurites of human DRG neurons. Our methods will enable the identification and characterisation of compounds being developed for the treatment of clinical mechanical hypersensitivity states.
    Original languageEnglish
    Pages (from-to)26-34
    Number of pages9
    JournalJournal of Neuroscience Methods
    Volume159
    Issue number1
    DOIs
    Publication statusPublished - 2007

    Keywords

    • Adult
    • Animals
    • Calcium Signaling
    • Cell Membrane
    • Cells, Cultured
    • Dendrites
    • Electrophysiology
    • Ganglia, Spinal
    • Humans
    • Male
    • Microscopy, Electron, Scanning
    • Microscopy, Fluorescence
    • Neurites
    • Neurons, Afferent
    • Physical Stimulation
    • Presynaptic Terminals
    • Rats
    • Rats, Wistar

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