The molecular mechanisms regulating intrinsic axon growth potential during development or following injury remain largely unknown, despite their vast importance. We have harnessed the genetic power of Drosophila to establish a neurite sprouting assay of primary cultured mushroom body (MB) neurons. Cultures are derived from brains in which the MB neurons are sparsely labelled (using the MARCM technique), thus allowing visualization and manipulation of single cells within the dense neuronal culture. Using this culturing system we can explore the sprouting characteristics of dissociated MB neurons – which mechanistically resembles axon regeneration after injury. By testing sprouting of WT and mutant MB neurons, this assay provides important insights about the intrinsic mechanisms of axon growth in both developmental and pathological contexts.
