The mushroom body of the insect brain is an important locus for olfactory information processing and associative learning. The present study investigated the biophysical properties of Kenyon cells, which form the mushroom body. Current- and voltage-clamp analyses were performed on cultured Kenyon cells from honeybees. Current-clamp analyses indicated that Kenyon cells did not spike spontaneously in vitro. However, spikes could be elicited by current injection in ~85% of the cells. Of the cells that produced spikes during a 1-s depolarizing current pulse, ~60 % exhibited repetitive spiking, whereas the remaining ~40% fired a single spike. Cells that spiked repetitively showed little frequency adaptation. However, spikes consistently became broader and smaller during repetitive activity. Voltage-clamp analyses characterized a fast transient Na⁺ current (I_Na), a delayed rectifier K⁺ current (I_K,V) and a fast transient K⁺ current (I_K,A). Using the neurosimulator SNNAP, a Hodgkin-Huxley type model was developed and used to investigate the roles of the different currents during spiking. The model led to the prediction of a slow transient outward current (I_K,ST) that was subsequently identified by re-evaluating the voltage-clamp data. Simulations indicated that the primary currents that underlie spiking are I_Na and I_K,V, whereas I_K,A and I_K,ST primarily determined the responsiveness of the model to stimuli such constant or oscillatory injections of current.