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      Cuticle preparation of Drosophila embryos and larvae.

      Methods in Molecular Biology (Clifton, N.j.)
      Animals, Developmental Biology, methods, Drosophila melanogaster, metabolism, physiology, Embryo, Nonmammalian, Epidermis, cytology, Genes, Insect, Genes, Reporter, Lac Operon, Larva, Models, Biological, Models, Genetic, Signal Transduction, Time Factors, Vitelline Membrane, beta-Galactosidase

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          Abstract

          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.

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