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We have constructed and characterized transgenic Drosophila lines with modified Na(+),K(+)-ATPase activity. Using a temperature dependent promoter from the hsp70 gene to drive expression of wild-type alpha subunit cDNA, we can conditionally rescue bang-sensitive paralysis and ouabain sensitivity of a Drosophila Na(+),K(+)-ATPase alpha subunit hypomorphic mutant, 2206. In contrast, a mutant alpha subunit (alpha(D369N)) leads to increased bang-sensitive paralysis and ouabain sensitivity. We can also generate temperature dependent phenotypes in wild-type Drosophila using the same hsp70 controlled alpha transgenes. Ouabain sensitivity was as expected, however, both bang sensitive paralysis or locomotor phenotypes became more severe regardless of the type of alpha subunit transgene. Using the Gal4-UAS system we have limited expression of alpha transgenes to cell types that normally express a particular Drosophila Na(+),K(+)-ATPase beta (Nervana) subunit isoform (Nrv1 or 2). The Nrv1-Gal4 driver results in lethality while the Nrv2-Gal4 driver shows reduced viability, locomotor function and uncontrolled wing beating. These transgenic lines will be useful for disrupting function in a broad range of cell types.

Antibodies recognizing horse radish peroxidase (HRP) stain neurons in Drosophila and other insects. We have used Western blots to analyze and characterize some of the anti-HRP reactive components from Drosophila melanogaster. Anti-HRP reactive components can be reproducibly detected during all developmental stages, although the pattern changes at different developmental times. In adults, there are at least 10 reproducibly stained components. Two of the bands, with molecular sizes of 42 and 80 kDa are likely to be the major contributors to neuronal anti-HRP staining in Drosophila. These components are enriched in adult fly heads. In contrast, many of the other anti-HRP reactive components in adults are enriched in abdomen and are present exclusively or at much higher levels in male flies. We have purified and characterized two of the male specific components with molecular sizes of 62 and 48 kDa. Partial N-terminal amino acid sequencing revealed that the 62 kDa protein is identical to a part of D. melanogaster carboxylesterase to EC 3.1.1.1) while he 48 kDa protein does not match any known sequences. Esterase-6 has previously been shown to be enriched in male accessory gland and consistent with this we show staining of this structure with anti-HRP antibodies.