The present study examined the involvement of N-methyl-d-aspartate (NMDA), α-amino-3-hydroxy-5-methyl-4-isoxazolpropionate/kainate (AMPA/KA) and dopamine receptors in the nucleus accumbens (ACB) in influencing reaction times of instrumental responses by the expectancy of reward. A simple reaction time task demanding conditioned lever release was used in which the upcoming reward magnitude was signalled in advance by discriminative cues. After training, in control rats with vehicle infusions (0.5 µL) into the ACB, reaction times of responses were significantly shorter to the discriminative cue predictive of high reward magnitude. Indirect stimulation of dopamine receptors in the ACB by d-amphetamine (20 µg/0.5 µL) decreased reaction times, impaired their guidance by cue-associated reward magnitudes and reduced the accuracy of task performance. Blockade of AMPA/KA receptors in the ACB by 6-cyano-7-nitroquino-xaline-2,3-dione (0.75 and 2.5 µg/0.5 µL) or NMDA receptors by d(−)-2-amino-5-phosphonopentanoic acid (5 µg/0.5 µL) produced a general increase in reaction times, but left guidance of reaction times by cue-associated reward magnitudes unaffected. Thus, stimulation of intra-ACB ionotropic glutamate receptors is critically involved in modulating the speed of instrumental responding to cues predictive for reward magnitude, but is not required for intact performance of previously learned instrumental behaviour.