Mechanism of the GAL4 Enhancer Trap with UAS-tau

Original images that appeared in Fig. 1 of:
Kei Ito, Heinz Sass, Joachim Urban, Alois Hofbauer and Stephan Schneuwly (1997)
GAL4-responsive UAS-tau as a tool for studying the anatomy and development of the Drosophila central nervous system
Cell and Tissue Research 290:1-10

The P-element enhancer-trap technique (O'Kane and Gehring, 1987) made it possible to easily screen thousands of strains that label various subsets of CNS cells. However, such strains were of limited anatomical use, as the original P-lacZ transposon labels only the nuclei.

An improved method uses yeast GAL4 that can activate any gene under control of the "upstream activation sequence" UAS (Fischer et al., 1988; Brand and Perrimon, 1993). Several cytoplasmic UAS-reporters are already available (Dunin and Brown, 1995; Ito et al., 1995; Yang et al., 1995; Yeh et al., 1995), but none of the constructs allow satisfactory labelling of the distal parts of axons and cell bodies simultaneously.

Here we tested a new UAS construct that uses the bovine tau gene. Tau is a microtubule-associated protein that moves from the cell body to the distal ends of nerve fibres via slow transportation, and should therefore provide an excellent marker for axonal staining. The distribution of the protein can be revealed with the commercially available monoclonal antibody against bovine Tau, which does not cross-react with intact Drosophila CNS.

Unlike reporters containing the large beta-galactosidase molecule, such as UAS-kinesin-lacZ and UAS-tau-lacZ(Callahan and Thomas, 1994; Ito et al., 1995), the small Tau protein should disturb the fine cytoskeleton structure less severely. The lack of lacZ in the construct also makes it possible to double-label GAL4- and lacZ-expressing cells.

Note:

References

• Brand AH, Perrimon N (1993) Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. Development 118:401-415
  
• Callahan CA, Thomas JB (1994) Tau-beta-galactosidase, an axon-targeted fusion protein. Proc Natn Acad Sci USA 91:5972-6
  
• Dunin BO, Brown NH (1995) Mammalian CD2 is an effective heterologous marker of the cell surface in Drosophila. Dev Biol 168:689-693
  
• Fischer JA, Giniger E, Maniatis T, Ptashne M (1988) GAL4 activates transcription in Drosophila. Nature 332:853-856
  
• Himmler A, Drechsel D, Kirschner MW, Martin DW (1989) Tau consists of a set of proteins with repeated C-terminal microtubule-binding domains and variable N-terminal domains. Mol Cell Biol 9:1381-1388
  
• Ito K, Urban J, Technau GM (1995) Distribution, classification, and development of Drosophila glial cells in the late embryonic and early larval ventral nerve cord. Roux's Arch Dev Biol 204:284-307
  
• O'Kane C, Gehring WJ (1987) Detection in situ of genomic regulatory elements in Drosophila. Proc Natn Acad Sci USA 84:9123-9127
  
• Yang MY, Armstrong JD, Vilinsky I, Strausfeld NJ, Kaiser K (1995) Subdivision of the Drosophila mushroom bodies by enhancer-trap expression patterns. Neuron 15:45-54
  
• Yeh E, Gustafson K, Boulianne GL (1995) Green fluorescent protein as a vital marker and reporter of gene expression in Drosophila. Proc Natl Acad Sci USA 92:7036-7040
  

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