The Notch Gene Product is a Glycoprotein Expressed on the Cell Surface of Both Epidermal and Neuronal Precursor Cells During Drosophila Development

 

Kristen M. Johansen, Richard G. Fehon and Spyros Artavanis-Tsakonas

 

ABSTRACT

 

The Notch locus of Drosophila melanogaster is one of a small number of zygotically acting "neurogenic" genes involved in the correct segregation of neural from epidermal lineages during embryogenesis as well as in other postembryonic developmental events. We have generated antibody probes against three regions of the Notch protein to study the expression of Notch and begin a biochemical characterization of the protein. Consistent with predictions based on DNA sequence data, here we gather evidence showing that Notch encodes a large, glycosylated surface protein with an apparent molecular weight of 300 kD: 1) all three antibodies detect Notch on Western blots as a high molecular weight, primarily full-length product; 2) immunoelectron microscopy localizes the Notch protein to the cell membrane; and 3) lentil lectin column binding demonstrates that the protein is glycosylated, indicative of its surface protein nature. In general, the distribution of the Notch protein coincides with that of the Notch transcripts determined previously by in situ hybridizations. Notch is expressed in a much wider range of tissue types than those disrupted in the neurogenic mutant, as determined by antibody localization. Early labelling in the blastoderm appears ubiquitous except for the pole cells, but as development proceeds some distinctive features emerge: stronger staining is seen within the germ band layer where neuroblast delamination occurs, and the developing embryonic nervous system shows pronounced axonal staining. In 3rd instar larvae Notch is expressed in imaginal discs and in the central nervous system. Based on these results, certain models for how Notch controls the neuroblast cell fate choice are eliminated. We discuss how Notch may function in this choice as well as in other lineage fate determinations.