Drosophila melanogaster has been a fruitful model for studying neurobiology, greatly increasing our understanding of neuronal fate determination, pathfinding, synapto-(genesis), ion channel function, neurotransmission (endocytosis and exo-cytosis), and synaptic plasticity. The primary strength of the Drosophila system is the ability to identify new molecules by systematic forward genetic screens; the only other model system where this approach is fully developed is Caenorhabditis elegans. The blind mutagenesis approach produces mutants in a process or pathway of interest without any bias based on preconceived ideas and is only limited by the speed and precision of the mutant screen (for review, see St. Johnston, 2002). A complementary approach, called reverse genetics, disrupts the activity of known proteins by the directed mutagenesis of genes (for review, see Adams and Sekelsky, 2002). In the past few years, much more rapid reverse genetic approaches have begun to be developed in Drosophila, including putative methods for targeted gene disruption (Rong and Golic, 2000; 2001; Bibikova et al., 2002) and RNA interference (RNAi; Kennerdell and Carthew, 1998; Misquitta and Patterson, 1999; Carthew et al., 2001; Schmid et al., 2002; Eaton et al., 2002; Allikian et al., 2002; Giordano et al., 2002). These new approaches allow more rapid tests of targeted gene function. Once mutants are generated, the fly nervous system is amenable to a wide range of cell biology approaches to elucidate the function of the gene product. A particularly powerful benefit of the Drosophila system is the ability to work with individually identified neurons, or small populations of neurons, derived through a known developmental lineage (Doe, 1992; Luer and Technau, 1992; for review, see Doe and Technau, 1993; Bossing and Techanau, 1994; Bossing et al, 1996a,b; Chu-LaGraff et al, 1991, 1993, 1995; Landgraf et al., 1997; Schmidt et al., 1997, 2000; Schmid et al., 1999). This chapter provides an overview of how an investigator can move from mutant to gene to gene product function using Drosophila as a genetic model system for cellular neurobiology. This chapter provides an outline of the resources and experimental approaches available to the Drosophila neurobiologist. Selected methods of particular interest are discussed in detail and protocols are provided.

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