We employed the retrograde neural tracing method using the pseudorabies virus bartha strain (PRV-Ba) to investigate the projection from accessory trigeminal nucleus (Acc5) or accessary facial nucleus (Acc7) to hypoglossal nucleus (HP). After injection of PRV-Ba into the bellies of the digastric muscle, the PRV-Ba containing neurones were observed in Acc5 of the cranial belly injected rats and Acc7 of caudal belly injected rats ipsilaterally, but not in HP. These results suggest that Acc5 and Acc7 may not project to HP and that movements of the digastric muscle are not related to cooperation with tongue movements during mastication.