Cellular mechanisms governing synaptic development in Drosophila melanogaster

Abstract

The neuromuscular connections of Drosophila are ideally suited for studying synaptic function and development. Hypotheses about cell recognition can be tested in a simple array of pre- and postsynaptic elements. Drosophila muscle fibers are multiply innervated by individually identifiable motoneurons. The neurons express several synaptic cotransmitters, including glutamate, proctolin, and octopamine, and are specialized by their synaptic morphology, neurotransmitters, and connectivity. During larval development the initial motoneuron endings grow extensively over the surface of the muscle fibers, and differentiate synaptic boutons of characteristic morphology. While considerable growth occurs postembryonically, the initial wiring of motoneurons to muscle fibers is accomplished during mid-to-late embryogenesis (stages 15-17). Efferent growth cones sample multiple muscle fibers with rapidly moving filopodia. Upon reaching their target muscle fibers, the growth cones rapidly differentiate into synaptic contacts whose morphology prefigures that of the larval junction. Mismatch experiments show that growth cones recognize specific muscle fibers, and can do so when the surrounding musculature is radically altered. However, when denied their normal targets, motoneurons can establish functional synapses on alternate muscle fibers. Blocking synaptic activity with either injected toxins or ion channel mutants does not derange synaptogenesis, but may influence the number of motor ending processes. The molecular mechanisms governing cellular recognition during synaptogenesis remain to be identified. However, several cell surface glycoproteins known to mediate cellular adhesion events in vitro are expressed by the developing synapses. Furthermore, enhancer detector lines have identified genes with expression restricted to small subsets of muscle fibers and/or motoneurons during the period of synaptogenesis. These observations suggest that in Drosophila a mechanism of target chemoaffinity may be involved in the genesis of stereotypic synaptic wiring.

A conditional tissue-specific transgene expression system using inducible GAL4

“…Filleted larval bodywall preparations were done as in ref. 28. Briefly, fillets were fixed in 4% paraformaldehyde for 40 min before blocking with 1% BSA and successive incubations with anti-even-skipped mAb 3C10 and AlexaFluor 568-labeled secondary Ab (Molecular Probes).…”

Drosophila Importin-α2 Is Involved in Synapse, Axon and Muscle Development

“…The importance of signaling pathways in establishing the normal pattern has been demonstrated previously by mutations in the toll/dorsal pathway and in dystroglycan and POMT1. The phenotypes of these mutants and others entail muscle duplications, absent muscles, branched muscles and errors in muscle insertion [57], [58], [59], [60], [61], [62]. All of these defects were observed in importin-α2 mutants and arose from the loss of Importin-α2 expression in the muscle (Figure 6 and Table 1).…”

‘Identify’ and ‘lock in’: molecular integration during synaptic target recognition

“…Several genetic screens have been designed to isolate mutations that cause synaptic targeting errors in specific sets of neurons [17,18]. In complementary studies that utilize the endogenous molecular expression patterns as entry points, the roles of genes expressed in specific subsets of neurons and/or their synaptic targets have been examined systematically [19][20][21]. However, the crucial question remains as to how these 'rare' molecules are coupled to a general machinery of synaptogenesis [5].…”