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      Notch signaling generates the “cut here line” on the cuticle of the puparium in Drosophila melanogaster

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          Summary

          During a molt or eclosion, insects shed their cuticle, an extracellular matrix made by underlying epidermal cells, by cleavage along a defined line. This means that the “cut here line” is pre-formed on the cuticle, and its formation is indispensable for insect life. Here, we show that the proper formation of the operculum ridge (OR), which is the “cut here line” on the puparium (pupal case) of Drosophila melanogaster, involves Notch signaling activation in the epidermal cells just beneath the future OR region (OR-forming cells). The inhibition of Notch signaling causes defects in eclosion due to failure in OR cleavage, the chitin organization and several cuticular proteins localization, glucose dehydrogenase (Gld) activity, and OR-forming cell shape. Our findings provide the first insight into the molecular basis of the structure and formation of the “cut here line” on the cuticle.

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          Highlights

          • Notch signaling in OR-forming cells activated by Ser is required for OR formation

          • Unique structure formation at the future OR region requires Notch signaling

          • Gld activation by Notch signaling is required for OR maturation in the pupal stage

          • Elongated OR-forming cell shape is regulated by Notch signaling

          Abstract

          Entomology; Evolutionary developmental biology

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          Most cited references29

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          A protein trap strategy to detect GFP-tagged proteins expressed from their endogenous loci in Drosophila.

          In Drosophila, enhancer trap strategies allow rapid access to expression patterns, molecular data, and mutations in trapped genes. However, they do not give any information at the protein level, e.g., about the protein subcellular localization. Using the green fluorescent protein (GFP) as a mobile artificial exon carried by a transposable P-element, we have developed a protein trap system. We screened for individual flies, in which GFP tags full-length endogenous proteins expressed from their endogenous locus, allowing us to observe their cellular and subcellular distribution. GFP fusions are targeted to virtually any compartment of the cell. In the case of insertions in previously known genes, we observe that the subcellular localization of the fusion protein corresponds to the described distribution of the endogenous protein. The artificial GFP exon does not disturb upstream and downstream splicing events. Many insertions correspond to genes not predicted by the Drosophila Genome Project. Our results show the feasibility of a protein trap in Drosophila. GFP reveals in real time the dynamics of protein's distribution in the whole, live organism and provides useful markers for a number of cellular structures and compartments.
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            A GAL4-driver line resource for Drosophila neurobiology.

            We established a collection of 7,000 transgenic lines of Drosophila melanogaster. Expression of GAL4 in each line is controlled by a different, defined fragment of genomic DNA that serves as a transcriptional enhancer. We used confocal microscopy of dissected nervous systems to determine the expression patterns driven by each fragment in the adult brain and ventral nerve cord. We present image data on 6,650 lines. Using both manual and machine-assisted annotation, we describe the expression patterns in the most useful lines. We illustrate the utility of these data for identifying novel neuronal cell types, revealing brain asymmetry, and describing the nature and extent of neuronal shape stereotypy. The GAL4 lines allow expression of exogenous genes in distinct, small subsets of the adult nervous system. The set of DNA fragments, each driving a documented expression pattern, will facilitate the generation of additional constructs for manipulating neuronal function. Copyright © 2012 The Authors. Published by Elsevier Inc. All rights reserved.
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              Staging the metamorphosis of Drosophila melanogaster.

              A sequence of 51 visible changes is described during the course of metamorphosis in Drosophila melanogaster, and a series of 24 convenient stages is defined for use in the experimental analysis and exploitation of this part of the insect life cycle. The duration of each stage is estimated and times are suggested for batch collections of symphasic animals.
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                Author and article information

                Contributors
                Journal
                iScience
                iScience
                iScience
                Elsevier
                2589-0042
                04 July 2023
                18 August 2023
                04 July 2023
                : 26
                : 8
                : 107279
                Affiliations
                [1 ]Department of Integrated Biosciences, Graduate School of Frontier Sciences, the University of Tokyo, Biosciences Building 501, 5-1-5 Kashiwanoha, Kashiwa-shi, Chiba 277-8562, Japan
                Author notes
                []Corresponding author tkojima@ 123456k.u-tokyo.ac.jp
                [2]

                Present address: Laboratory for Extracellular Morphogenesis, Graduate School of Science, Chiba University, 1–33 Yayoi-cho, Inage-ku, Chiba-shi, Chiba 263–8522, Japan

                [3]

                Lead contact

                Article
                S2589-0042(23)01356-1 107279
                10.1016/j.isci.2023.107279
                10387574
                e0ea392a-c862-41f3-9bde-3f798e54fb3f
                © 2023 The Author(s)

                This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

                History
                : 17 January 2023
                : 25 May 2023
                : 30 June 2023
                Categories
                Article

                entomology,evolutionary developmental biology
                entomology, evolutionary developmental biology

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