title: Amplification and expression of a salivary gland DNA puff gene in the prothoracic gland of Bradysia hygida (Diptera: Sciaridae).
abstract: The DNA puff BhC4-1 gene, located in DNA puff C4 of Bradysiahygida, is amplified and expressed in the salivary gland at the end of the fourth larval instar as a late response to the increase in 20-hydroxyecdysone titer that triggers metamorphosis. Functional studies revealed that the mechanisms that regulate BhC4-1 expression in the salivary gland are conserved in transgenic Drosophila. These studies also led to the identification of a cis-regulatory module that drives developmentally regulated expression of BhC4-1-lacZ in the prothoracic gland cells of the ring gland, a compound organ which in Drosophila results from the fusion of the prothoracic glands, the corpus allatum and the corpus cardiacum. Here we have investigated the occurrence of BhC4-1 expression in B. hygida prothoracic glands. We report the identification of the B. hygida prothoracic gland and demonstrate that it releases ecdysone. Using RT-qPCR, western blots and immunolocalization experiments, we demonstrate that the BhC4-1 mRNA and the BhC4-1 protein are both expressed in the B. hygida prothoracic glands at the same time that DNA puff C4 is formed in the salivary gland. We also show that BhC4-1 is concomitantly amplified 4.8-fold in the prothoracic gland and 23-fold in the salivary gland. Our results reveal the occurrence of stage specific expression of a DNA puff gene in the prothoracic glands of B. hygida, and extend previous studies that have shown that DNA puff genes expression is not restricted to the salivary gland. In addition, the description of stage specific gene amplification in the prothoracic glands of B. hygida constitutes the first demonstration that gene amplification in Diptera might occur concomitantly in two different tissues in the same developmental stage.
authors: Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP CEP 14040-903, Brazil. Electronic address: julianacandido@facica.edu.br.
authors: Juliana Aparecida
authors: JA
authors: Candido-Silva
authors: Departamento de Biologia Celular e Molecular e Bioagentes Patogênicos, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP CEP 14049-900, Brazil. Electronic address: maiarocabral@hotmail.com.
authors: Maiaro Cabral Rosa
authors: MC
authors: Machado
authors: Departamento de Biologia Celular e Molecular e Bioagentes Patogênicos, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP CEP 14049-900, Brazil. Electronic address: klaus@fmrp.usp.br.
authors: Klaus Hartmann
authors: KH
authors: Hartfelder
authors: Departamento de Biologia Celular e Molecular e Bioagentes Patogênicos, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP CEP 14049-900, Brazil. Electronic address: jcdalmei@fmrp.usp.br.
authors: Jorge Cury
authors: JC
authors: de Almeida
authors: Departamento de Biologia Celular e Molecular e Bioagentes Patogênicos, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP CEP 14049-900, Brazil. Electronic address: mlplarso@fmrp.usp.br.
authors: Maria Luisa
authors: ML
authors: Paçó-Larson
authors: Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP CEP 14040-903, Brazil. Electronic address: namonesi@fcfrp.usp.br.
authors: Nadia
authors: N
authors: Monesi
mesh_terms: D000818:Animals; D004175:Diptera / Q000235:genetics* / Q000254:growth & development* / Q000378:metabolism; D004441:Ecdysterone / Q000378:metabolism*; D004702:Endocrine Glands / Q000378:metabolism; D005784:Gene Amplification; D017344:Genes, Insect; D019476:Insect Proteins / Q000235:genetics* / Q000378:metabolism; D007814:Larva / Q000254:growth & development / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012471:Salivary Proteins and Peptides / Q000235:genetics* / Q000378:metabolism
chemical_list: D019476:Insect Proteins; D012333:RNA, Messenger; D012471:Salivary Proteins and Peptides; C095605:puff C4 protein, Bradysia hygida; D004441:Ecdysterone
keywords: DNA puff genes; Gene amplification; Prothoracic glands; Sciaridae