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Searching for the molecular mediators of motor learning in Drosophila

Torque learning in tethered Drosophila is an example of operant self-learning that can be classified as an instance of motor learning. Vocal learning in birds and humans is also a form of motor learning and requires protein kinase C (PKC) and the transcription factors of the FoxP family. Operant learning in the marine snail Aplysia also requires PKC and is independent of adenylyl cyclase type I, important for many other forms of learning. Also in Drosophila, motor learning is independent of the rutabaga type I cyclase and requires the atypical PKC (aPKC) and FoxP, suggesting that motor learning relies on a conserved molecular pathway. To discover additional components of this conserved pathway, we use a two-pronged approach: we not only screen candidate genes, using CRISPR/Cas9-mediated knock-outs, for defects in motor learning, we also create transgenic flies with marked components of the pathway. First knock-out results show that N-cadherin appears not to be involved in motor learning in Drosophila. Further candidates are neurexin-1, the neuroligins and the RS6 Kinase II ignorant. To identify aPKC interacting proteins, we have generated transgenic flies expressing a biotin ligase fused to the aPKC protein. This fusion protein will biotinylate targets in close proximity to aPKC, such that they can be isolated using streptavidin beads for later analysis.