Characterizing the embryonic development of B. hygida (Diptera: Sciaridae) following enzymatic treatment to permeabilize the serosal cuticle
| Autor: | João Vitor Cardoso Uliana, Luciano Antonio Digiampietri, Nadia Monesi, Luiz Paulo Andrioli, James Castelli-Gair Hombría, Guilherme T.P. Brancini |
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| Přispěvatelé: | Fundação de Amparo à Pesquisa do Estado de São Paulo |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Embryology Cuticle Embryonic Development Chitin 03 medical and health sciences chemistry.chemical_compound Melanogaster Serosal cuticle Animals Sciaridae biology Diptera fungi Embryogenesis Chitin synthase Embryo biology.organism_classification Biological Evolution Cell biology Drosophila melanogaster 030104 developmental biology chemistry biology.protein DIPTERA Transcriptome Developmental Biology |
| Zdroj: | Repositório Institucional da USP (Biblioteca Digital da Produção Intelectual) Universidade de São Paulo (USP) instacron:USP Digital.CSIC: Repositorio Institucional del CSIC Consejo Superior de Investigaciones Científicas (CSIC) Digital.CSIC. Repositorio Institucional del CSIC instname |
| Popis: | Understanding the evolution of the developmental programs active during dipteran embryogenesis depends on comparative studies. As a counterpoint to the intensively investigated and highly derived cyclorrhaphan flies that include the model organism Drosophila melanogaster, we are studying the basal Diptera Bradysia hygida, a member of the Sciaridae family that is amenable to laboratory cultivation. Here we describe the B. hygida embryogenesis, which lasts 9 days at 22 °C. The use of standard fixation D. melanogaster protocols resulted in embryos refractory to DAPI staining and to overcome this, a new enzyme-based method was developed. Calcofluor-White staining of enzimatically-treated embryos revealed that this method removes chitin from the serosal cuticle surrounding the B. hygida embryo. Chitin is one of the main components of serosal cuticles and searches in a B. hygida embryonic transcriptome database revealed conservation of the chitin synthesis pathway, further supporting the occurrence of chitin biosynthesis in B. hygida embryos. Combining the enzymatic treatment protocol with the use of both DIC and fluorescence microscopy allowed the first complete description of the B. hygida embryogenesis. Our results constitute an important step towards the understanding of early development of a basal Diptera and pave the way for future evo-devo studies. Work in N. Monesi's and L.P. Andrioli's laboratories was supported by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) (grant numbers 2016/25325-8 and 2014/14318-5, respectively). JVCU was the recipient of a FAPESP undergraduate scholarship (grant number 2016/00412-5). We also thank FAPESP for a Ph.D. scholarship to GTPB (grant number 2015/24305-0). |
| Databáze: | OpenAIRE |
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