The Wnt family of lipoproteins regulates several aspects of the development of the nervous system. Recently, we reported that Wnt‐3a enhances the proliferation of midbrain dopaminergic precursors and that Wnt‐5a promotes their differentiation into dopaminergic neurones. Here we report the purification of hemagglutinin‐tagged Wnt‐5a using a three‐step purification method similar to that previously described for Wnt‐3a. Haemagglutinin‐tagged Wnt‐5a was biologically active and induced the differentiation of immature primary midbrain precursors into tyrosine hydroxylase‐positive dopaminergic neurones. Using a substantia nigra‐derived dopaminergic cell line (SN4741), we found that Wnt‐5a, unlike Wnt‐3a, did not promote β‐catenin phosphorylation or stabilization. However, both Wnt‐5a and Wnt‐3a activated dishevelled, as assessed by a phosphorylation‐dependent mobility shift. Moreover, the activity of Wnt‐5a on dishevelled was blocked by pre‐treatment with acyl protein thioesterase‐1, indicating that palmitoylation of Wnt‐5a is necessary for its function. Thus, our results suggest that Wnt‐3a and Wnt‐5a, respectively, activate canonical and non‐canonical Wnt signalling pathways in ventral midbrain dopaminergic cells. Furthermore, we identify dishevelled as a key player in transducing both Wnt canonical and non‐canonical signals in dopaminergic cells.