In vivo modification of Na+,K+-ATPase activity in Drosophila

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 a 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 a transgenes. Ouabain sensitivity was as expected, however, both bang sensitive paralysis or locomotor phenotypes became more severe regardless of the type of a subunit transgene. Using the Gal4-UAS system we have limited expression of a 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. (C) 2001 Elsevier Science Inc. All rights reserved.