Cuticle preparation of Drosophila embryos and larvae.
The Drosophila embryonic ventral epidermis has served as a unique tissue for the genetic analysis of patterning. Two types of epidermal cells are easily distinguished: those that secrete short, thick hair-like structures called denticles and cells that only secrete smooth cuticle. Denticle-secreting cells form segmentally repeated belts. Within each belt, six types of denticles can be recognized according to size, shape, and orientation (types 1-6). They are arranged in a stereotypical manner within each denticle belt. This pattern results from the spatially organized activation of several signaling pathways during embryogenesis. Cuticle patterns therefore provide a sensitive readout of signaling activity and other patterning mechanisms. Here, I describe methods of preparation and analysis of cuticles from 1st instar larvae as well as from 3rd instar larvae. In addition, a protocol to simultaneously analyze cuticles and beta-galactosidase activity of embryos expressing lacZ reporter genes is presented.
Alexandre
Cyrille
C
National Institute for Medical Research, The Ridgeway Mill Hill, London, UK.
D000818:Animals; D015509:Developmental Biology / Q000379:methods; D004331:Drosophila melanogaster / Q000378:metabolism* / Q000502:physiology*; D004625:Embryo, Nonmammalian; D000078404:Epidermal Cells; D004817:Epidermis / Q000378:metabolism*; D017344:Genes, Insect; D017930:Genes, Reporter; D007763:Lac Operon; D007814:Larva / Q000378:metabolism; D008954:Models, Biological; D008957:Models, Genetic; D015398:Signal Transduction; D013997:Time Factors; D014817:Vitelline Membrane / Q000378:metabolism; D001616:beta-Galactosidase / Q000378:metabolism
D001616:beta-Galactosidase