Immunological characterization of K+ channel components from the Shaker locus and differential distribution of splicing variants in Drosophila.

Antibodies were raised to three portions of the predicted sequences of Shaker, a gene that encodes a family of K+ channel components that are produced by the alternative splicing of transcripts. On immunoblots, the protein products appear to be 65,000-85,000 daltons in size. No smaller products were detected. Immunocytochemistry has revealed a nonuniform distribution of Shaker products in the brain of the adult fly. By comparing antisera directed against regions shared by all the splicing variants to antisera that are directed against one particular group of splicing variants, we have determined that there is a differential distribution of that group of variants. Thus, the alternative splicing of Shaker transcripts appears to produce different subtypes of A-channels in different tissues.

Searching for pattern and mutation in the Drosophila genome with a P-lacZ vector.

A P-element vector has been constructed and used to generate lines of flies with single autosomal P-element insertions. The lines were analyzed in two ways: (1) the identification of cis-acting patterning information within the Drosophila genome, as revealed by a lacZ reporter gene within the P element, and (2) the isolation of lethal mutations. We examined 3768 independent lines for the expression of lacZ in embryos and looked among these lines for lethal mutations affecting embryonic neurogenesis. This type of screen appears to be an effective way to find new loci that may play a role in the development of the Drosophila nervous system.

Neurogenesis of the peripheral nervous system in Drosophila embryos: DNA replication patterns and cell lineages.

Cell lineages that give rise to the PNS were studied using the thymidine analog 5-bromo-2'-deoxyuridine (BrdU) to visualized DNA replication immunocytochemically. The precursors of the PNS in the body segments of Drosophila embryos replicate their DNA in a spatially and temporally stereotyped pattern. The sequence of DNA replication within developing sensory organs suggests particular lineage relationships of the cells that constitute a sensory organ, i.e., neuron and associated support cells. In embryos that are mutant for the achaete-scute complex or daughterless, in which most or all of the PNS is missing, no BrdU-labeled cells were found in the appropriate regions, suggesting that these PNS precursors either do not form or fail to replicate. Thus, the BrdU technique allows determination as to whether a mutation affects the PNS precursors or terminal differentiation.