One way to expand the existing range of anti‐migraine drugs seems to be the search for pharmacological agents with anti‐cephalalgic properties among medicines approved for clinical indications other than migraine. Numerous experimental and clinical data imply that selective serotonin 5‐HT3 receptor antagonists can be considered as potential anti‐migraine agents. Therefore, the objective of our work was to examine the impact of selective 5‐HT3 receptor blockade with granisetron on migraine‐related nociceptive transmission within the spinal trigeminal nucleus (STN) and the ventroposteromedial nucleus of the thalamus (VPM). Using an electrophysiological model of trigemino‐durovascular nociception in anaesthetised male Wistar rats, we evaluated the effects of intravenous administration of granisetron on ongoing firing and dural electrical stimulation‐evoked responses of the spinal trigeminal and thalamic cells. Granisetron did not substantially affect responses of the STN and VPM neurons to electrical stimulation of the dura mater as well as did not cause steady changes in ongoing firing of the spinal trigeminal cells. The results obtained argue against the use of 5‐HT3 receptor antagonists for treating migraine. These data also lead to the conclusion that in the absence of sustained sensitisation of neurons along the trigemino‐thalamo‐cortical pathway the role of 5‐HT3 receptor‐dependent mechanisms in serotonergic modulation of trigeminovascular nociceptive transmission can hardly be considered crucial.