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Calreticulin expression levels and endoplasmic reticulum during late oogenesis and early embryogenesis of Rhodnius prolixus Stahl

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Abstract

This study reports the cloning, expression analysis and localization of calreticulin (CRT) in the endoplasmic reticulum (ER) during late oogenesis and early embryogenesis of the insect Rhodnius prolixus. CRT was cloned and sequenced from cDNA extracted from unfertilized eggs. Real-time PCR showed that CRT expression remains at lower levels during late oogenesis when compared to vitellogenic oocytes or day 0 laid fertilized eggs. Immunofluorescence microscopy showed that this protein is located in the periphery of the egg, in a differential peripheral ooplasm surrounding the yolk-rich internal ooplasm, only identified by transmission electron microscopy (TEM) of thin sections. Using immunogold electron microscopy, the ER ultrastructure (CRT labeled) was identified in the peripheral ooplasm as dispersed lamellae, randomly distributed in the peripheral ooplasm. No massive alterations of ER ultrastructure were found before or right after (30 min) fertilization, but an increase in CRT expression levels and assembly of typical rough ER (parallel cisternae with associated ribosomes) were observed 18–24 h after oviposition. The lack of ER assembly at fertilization and the later formation of rough ER together with the increase in CRT expression levels, suggest that the major functions of ER might be of great importance during the early events of development. The possible involvement of ER in the early steps of embryogenesis will be discussed.

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Acknowledgments

The authors thank Francisco G. Nobrega and Kildare Miranda. This study was supported by grants from Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq/Brazil), Fundação Carlos Chagas Filho de Amparo à Pesquisa no Estado do Rio de Janeiro (FAPERJ), Programa Pensa Rio, Programa de Pesquisadores Emergentes, Programa de Apoio ao Desenvolvimento Científico e Tecnológico (PADCT), and FINEP/PRONEX/FUJB no. 76.97.1000.000 (to EAM).

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Authors and Affiliations

  1. Laboratório de Entomologia Médica, Instituto de Biofísica Carlos Chagas Filho (IBCCF), Universidade Federal do Rio de Janeiro(UFRJ), Rio de Janeiro, Brazil

    Isabela B. Ramos & Ednildo A. Machado

  2. Laboratório de Ultraestrutura Celular Hertha Meyer, Instituto de Biofísica Carlos Chagas Filho (IBCCF), Universidade Federal do Rio de Janeiro(UFRJ), Rio de Janeiro, Brazil

    Isabela B. Ramos & Wanderley de Souza

  3. Laboratrio de Biologia Molecular e Celular de Fungos, UNIVAP, São José dos Campos, Brazil

    Claudia B. L. Campos

  4. Laboratório de Bioquímica de Artrópodos Hematófagos, Instituto de Bioquímica Médica (IBqM), Universidade Federal do Rio de Janeiro(UFRJ), Rio de Janeiro, Brazil

    Marcos H. F. Sorgine

  5. Instituto Nacional de Metrologia, Normalização e Qualidade Industrial—Inmetro, Rio de Janeiro, Brazil

    Wanderley de Souza

  6. Centro de Ciências da Saúde (CCS), Universidade Federal do Rio de Janeiro (UFRJ), Cidade Universitária—Ilha do Fundão, Rio de Janeiro, RJ, 21941-590, Brazil

    Ednildo A. Machado

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  1. Isabela B. Ramos
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  2. Claudia B. L. Campos
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  3. Marcos H. F. Sorgine
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  4. Wanderley de Souza
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  5. Ednildo A. Machado
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Correspondence to Ednildo A. Machado.

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Ramos, I.B., Campos, C.B.L., Sorgine, M.H.F. et al. Calreticulin expression levels and endoplasmic reticulum during late oogenesis and early embryogenesis of Rhodnius prolixus Stahl. Mol Biol Rep 38, 1757–1767 (2011). https://doi.org/10.1007/s11033-010-0290-0

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  • DOI: https://doi.org/10.1007/s11033-010-0290-0

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