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Developmental ecdysteroid titers and DNA puffs in larvae of two sciarid species, Rhynchosciara americana and Rhynchosciara milleri (Diptera: Sciaridae)

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Abstract

Ecdysteroid titers, developmental landmarks and the presence of prominent amplifying regions (DNA puffs) have been compared during late larval to pupal development in four groups of Rhynchosciara americana larvae and in R. americana and Rhynchosciara milleri. Three prominent DNA puffs (B2, C3 and C8) expand and regress sequentially on the rising phase of the 20-hydroxyecdysone (20E) titer in R. americana as a firm, cellular cocoon is being constructed. A sharp rise in 20E coincides with the regression of these puffs. The shape of the 20E curve is similar in R. milleri, a species that does not construct a massive cocoon, but the behavior of certain DNA puffs and their temporal relationship to the curve differs. Regions corresponding to B2 and C3 can be identified in R. milleri by banding pattern similarity with R. americana chromosomes and, in the case of B2, by hybridization to an R. americana probe. A B2 puff appears in R. milleri as the 20E titer rises but remains small in all gland regions. A puff similar to the R. americana C3 puff occurs in posterior gland cells of R. milleri (C3Rm) after the B2 puff, but this site did not hybridize to R. americana C3 probes. C3Rm incorporated 3H-thymidine above background, but showed less post-puff DNA accumulation than C3 of R. americana. R. americana C8 probes hybridized to a more distal region of the R. milleri C chromosome that did not appear to amplify or form a large puff. These differences can be related to developmental differences, in particular differences in cocoon construction between the two species.

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Acknowledgments

We thank the University of São Paulo, Department of Genetics and Evolutionary Biology for support during the time when many of the experiments were carried out. The ecdysone antibody was generated and kindly provided by the Gilbert lab (UNC Chapel Hill.) We also thank Michele Santos Machado for assistance in the collection of data for R. milleri and Jason Axford for his help in resizing the graphs. The Fundação de Amparo e Pesquisa do Estado de São Paulo (FAPESP) provided financial support through a grant to AJS.

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Authors and Affiliations

  1. Departamento de Biologia Estrutural, Molecular e Genética, Universidade Estadual de Ponta Grossa, Ponta Grossa, Paraná, Brazil

    M. A. M. Soares

  2. Departamento de Biologia, Celular e Molecular e de Bioagentes Patogênicos, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brazil

    K. Hartfelder

  3. Departamento Acadêmico de Química e Biologia, Universidade Tecnológica Federal do Paraná – Sede Ecoville, Rua Deputado Heitor de Alencar Furtado, Curitiba, PR, Brazil

    J. M. Tesserolli de Souza

  4. School of BioSciences, University of Melbourne, 30 Flemington Rd., Parkville, VIC, 3052, Australia

    A. J. Stocker

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Soares, M.A.M., Hartfelder, K., Tesserolli de Souza, J.M. et al. Developmental ecdysteroid titers and DNA puffs in larvae of two sciarid species, Rhynchosciara americana and Rhynchosciara milleri (Diptera: Sciaridae). Genetica 143, 597–612 (2015). https://doi.org/10.1007/s10709-015-9859-9

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