Continuous electrophysiological measurements of changes in cell volume of motoneurons in the isolated frog spinal cord

Abstract

One type of ion-sensitive micro-electrode (K+ ligand Corning 477317) is sensitive to large quaternary ammonium ions such as choline or tetramethylammonium (TMA+). We have now used such electrodes for continuous electrophysiological measurements of changes in cell volume of motoneurons in the isolated frog spinal cord. The electrodes were double-barrelled with tip diameters of 1 micron. The reference barrel was filled with 100 mM choline or 100 mM TMA+ in 1 M Mg2+-acetate, the sensitive barrel contained the Corning K+ ligand. After the impalement of a motoneuron, choline or TMA+ diffused into the cell and about 1 h later, a steady-state concentration of these ions in the range of 10-20 mM was reached. Following this period, the motoneurons were activated by repetitive electrical stimulation or by application of amino acids via the bathing solution. All these stimuli led to a transient rise of the intracellular concentrations of choline or TMA+ (indicating a cell shrinkage of 3-10% difference to control volume).