1. Intracellular records were obtained from giant interneurones in the isolated spinal cord of the sea lamprey. The cells had a mean resting potential of about 75 mV and action potentials with overshoots of about 35 mV. Their input resistances, measured by passing polarizing currents through the recording pipette, were in the range 3-7 MOmega.2. Iontophoretic ejection of gamma-aminobutyric acid (GABA) from a micropipette placed near the surface of a cell resulted in a slight hyperpolarization, accompanied by a marked reduction in input resistance. The reversal point for the potential change was about 5 mV greater than the resting membrane potential.3. Iontophoretic application of L-glutamate to the cells produced a depolarization with a decrease in input resistance much smaller than that accompanying a GABA potential of similar amplitude. The action potential amplitude was reduced by L-glutamate application. The reversal potential could not be determined accurately but appeared to be near zero membrane potential.4. Glutamate application produced, in addition, a burst of inhibitory synaptic potentials in the cell, presumably by depolarizing either inhibitory presynaptic nerve terminals or nearby inhibitory cell bodies.5. Acetylcholine (ACh) produced no detectable change in membrane resistance or potential.6. Application of the three drugs to first-order sensory cells in the spinal cord had no effect on their membrane properties.