Molecular Biology Reports

, Volume 38, Issue 3, pp 1757–1767 | Cite as

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

  • Isabela B. Ramos
  • Claudia B. L. Campos
  • Marcos H. F. Sorgine
  • Wanderley de Souza
  • Ednildo A. Machado
Article
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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.

Keywords

Calreticulin Eggs Endoplasmic reticulum Rhodnius prolixus 
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Notes

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).

References

  1. 1.
    Kunkel JG, Nordin JH (1985) Yolk proteins: comprehensive insect physiology, biochemistry and pharmacology. Pergamon Press, OxfordGoogle Scholar
  2. 2.
    Raikhel AS, Dhadialla TS (1992) Accumulation of yolk proteins in insect oocytes. Annu Rev Entomol 37:217–251CrossRefPubMedGoogle Scholar
  3. 3.
    Kerkut GG, Gilbert LG (1985) Comprehensive insect physiology, biochemistry and pharmacology. Pergamon Press, OxfordGoogle Scholar
  4. 4.
    Page AW, Orr-Weaver TL (1997) Stopping and starting the meiotic cell cycle. Curr Opin Genet Dev 7:23–31CrossRefPubMedGoogle Scholar
  5. 5.
    Whitaker M (2006) Calcium at fertilization and in early development. Physiol Rev 86:25–88CrossRefPubMedGoogle Scholar
  6. 6.
    Carafoli E, Santella L, Branca D, Brini M (2001) Generation, control, and processing of cellular calcium signals. Crit Rev Biochem Mol Biol 36:107–260CrossRefPubMedGoogle Scholar
  7. 7.
    Terasaki M, Jaffe LA (1991) Organization of the sea urchin egg endoplasmic reticulum and its reorganization at fertilization. J Cell Biol 114:929–940CrossRefPubMedGoogle Scholar
  8. 8.
    Terasaki M, Jaffe LA, Hunnicutt GR, Hammer JA 3rd (1996) Structural change of the endoplasmic reticulum during fertilization: evidence for loss of membrane continuity using the green fluorescent protein. Dev Biol 179:320–328CrossRefPubMedGoogle Scholar
  9. 9.
    Bobinnec Y, Marcaillou C, Morin X, Debec A (2003) Dynamics of the endoplasmic reticulum during early development of Drosophila melanogaster. Cell Motil Cytoskeleton 54:217–225CrossRefPubMedGoogle Scholar
  10. 10.
    McCauliffe DP, Yang YS, Wilson J, Sontheimer RD, Capra JD (1992) The 5’-flanking region of the human calreticulin gene shares homology with the human GRP78, GRP94, and protein disulfide isomerase promoters. J Biol Chem 267:2557–2562PubMedGoogle Scholar
  11. 11.
    Gamo S, Dodo K, Matakatsu H, Tanaka Y (1998) Molecular genetical analysis of Drosophila ether sensitive mutants. Toxicol Lett 100–101:329–337CrossRefPubMedGoogle Scholar
  12. 12.
    Gamo S, Tomida J, Dodo K, Keyakidani D, Matakatsu H et al (2003) Calreticulin mediates anesthetic sensitivity in Drosophila melanogaster. Anesthesiology 99:867–875CrossRefPubMedGoogle Scholar
  13. 13.
    Michalak M, Groenendyk J, Szabo E, Gold LI, Opas M (2009) Calreticulin, a multi-process calcium-buffering chaperone of the endoplasmic reticulum. Biochem J 417:651–666CrossRefPubMedGoogle Scholar
  14. 14.
    Michalak M, Mariani P, Opas M (1998) Calreticulin, a multifunctional Ca2+ binding chaperone of the endoplasmic reticulum. Biochem Cell Biol 76:779–785CrossRefPubMedGoogle Scholar
  15. 15.
    Michalak M, Corbett EF, Mesaeli N, Nakamura K, Opas M (1999) Calreticulin: one protein, one gene, many functions. Biochem J 344(Pt 2):281–292CrossRefPubMedGoogle Scholar
  16. 16.
    Stoltzfus JR, Horton WJ, Grotewiel MS (2003) Odor-guided behavior in Drosophila requires calreticulin. J Comp Physiol A Neuroethol Sens Neural Behav Physiol 189:471–483CrossRefPubMedGoogle Scholar
  17. 17.
    Zhang L, Wu G, Tate CG, Lookene A, Olivecrona G (2003) Calreticulin promotes folding/dimerization of human lipoprotein lipase expressed in insect cells (sf21). J Biol Chem 278:29344–29351CrossRefPubMedGoogle Scholar
  18. 18.
    Asgari S, Schmidt O (2003) Is cell surface calreticulin involved in phagocytosis by insect hemocytes? J Insect Physiol 49:545–550CrossRefPubMedGoogle Scholar
  19. 19.
    Johnson S, Michalak M, Opas M, Eggleton P (2001) The ins and outs of calreticulin: from the ER lumen to the extracellular space. Trends Cell Biol 11:122–129CrossRefPubMedGoogle Scholar
  20. 20.
    Garcia ES, Macarini JD, Garcia ML, Ubatuba FB (1975) Feeding of Rhodnius prolixus in the laboratory. Anais da Academia Brasileira de Ciencias 47:537–545PubMedGoogle Scholar
  21. 21.
    Fialho E, Silveira AB, Masuda H, Silva-Neto MA (2002) Oocyte fertilization triggers acid phosphatase activity during Rhodnius prolixus embryogenesis. Insect Biochem Mol Biol 32:871–880CrossRefPubMedGoogle Scholar
  22. 22.
    Fialho E, Nakamura A, Juliano L, Masuda H, Silva-Neto MA (2005) Cathepsin D-mediated yolk protein degradation is blocked by acid phosphatase inhibitors. Arch Biochem Biophys 436:246–253CrossRefPubMedGoogle Scholar
  23. 23.
    Heming BaH E (1994) Development of the germ cels and reproductive primordia in male and female embryos of Rhodnius prolixus Stahl (hemiptera: reduviidae). Can J Zool 72:1100–1119CrossRefGoogle Scholar
  24. 24.
    Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25:402–408CrossRefPubMedGoogle Scholar
  25. 25.
    Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275PubMedGoogle Scholar
  26. 26.
    Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685CrossRefPubMedGoogle Scholar
  27. 27.
    Berryman MA, Rodewald RD (1990) An enhanced method for post-embedding immunocytochemical staining which preserves cell membranes. J Histochem Cytochem 38:159–170PubMedGoogle Scholar
  28. 28.
    Tutunco L, Stein P, Ord TS, Jorgez CJ, Willians CJ (2004) Calreticulin on the mouse egg surface mediates transmembrane signaling linked to cell cycle resumption. Dev Biol 270:246–260CrossRefGoogle Scholar
  29. 29.
    Gilbert SF (2000) Developmental Biology. Sinauer Associates, Inc, SunderlandGoogle Scholar
  30. 30.
    Kelly GH, Huebner E (1989) Embryonic development of the hemipteran insect Rhodnius prolixus. J Morphol 199:175–196CrossRefGoogle Scholar
  31. 31.
    Schier AF (2007) The maternal-zygotic transition: death and birth of RNAs. Science 316:406–407CrossRefPubMedGoogle Scholar
  32. 32.
    Stitzel ML, Seydoux G (2007) Regulation of the oocyte-to-zygote transition. Science 316:407–408CrossRefPubMedGoogle Scholar
  33. 33.
    Sardet C, Prodon F, Dumollard R, Chang P, Chenevert J (2002) Structure and function of the egg cortex from oogenesis through fertilization. Dev Biol 241:1–23CrossRefPubMedGoogle Scholar
  34. 34.
    Heifetz Y, Yu J, Wolfner MF (2001) Ovulation triggers activation of Drosophila oocytes. Dev Biol 234:416–424CrossRefPubMedGoogle Scholar
  35. 35.
    Horner VL, Czank A, Jang JK, Singh N, Williams BC et al (2006) The Drosophila calcipressin sarah is required for several aspects of egg activation. Curr Biol 16:1441–1446CrossRefPubMedGoogle Scholar
  36. 36.
    Takeo S, Tsuda M, Akahori S, Matsuo T, Aigaki T (2006) The calcineurin regulator sra plays an essential role in female meiosis in Drosophila. Curr Biol 16:1435–1440CrossRefPubMedGoogle Scholar
  37. 37.
    Horner VL, Wolfner MF (2008) Mechanical stimulation by osmotic and hydrostatic pressure activates Drosophila oocytes in vitro in a calcium-dependent manner. Dev Biol 316:100–109CrossRefPubMedGoogle Scholar
  38. 38.
    Ramos IB, Miranda K, de Souza W, Oliveira DM, Lima AP et al (2007) Calcium-regulated fusion of yolk granules is important for yolk degradation during early embryogenesis of Rhodnius prolixus Stahl. J Exp Biol 210:138–148CrossRefPubMedGoogle Scholar
  39. 39.
    Schroder R (2003) The genes orthodenticle and hunchback substitute for bicoid in the beetle Tribolium. Nature 422:621–625CrossRefPubMedGoogle Scholar
  40. 40.
    Liu PZ, Kaufman TC (2005) Short and long germ segmentation: unanswered questions in the evolution of a developmental mode. Evol Dev 7:629–646CrossRefPubMedGoogle Scholar
  41. 41.
    Edgar BA, Schubiger G (1986) Parameters controlling transcriptional activation during early Drosophila development. Cell 44:871–877CrossRefPubMedGoogle Scholar
  42. 42.
    Parry H, McDougall A, Whitaker M (2005) Microdomains bounded by endoplasmic reticulum segregate cell cycle calcium transients in syncytial Drosophila embryos. J Cell Biol 171:47–59CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Isabela B. Ramos
    • 1
    • 2
  • Claudia B. L. Campos
    • 3
  • Marcos H. F. Sorgine
    • 4
  • Wanderley de Souza
    • 2
    • 5
  • Ednildo A. Machado
    • 1
    • 6
  1. 1.Laboratório de Entomologia Médica, Instituto de Biofísica Carlos Chagas Filho (IBCCF)Universidade Federal do Rio de Janeiro(UFRJ)Rio de JaneiroBrazil
  2. 2.Laboratório de Ultraestrutura Celular Hertha Meyer, Instituto de Biofísica Carlos Chagas Filho (IBCCF)Universidade Federal do Rio de Janeiro(UFRJ)Rio de JaneiroBrazil
  3. 3.Laboratrio de Biologia Molecular e Celular de FungosUNIVAPSão José dos CamposBrazil
  4. 4.Laboratório de Bioquímica de Artrópodos Hematófagos, Instituto de Bioquímica Médica (IBqM)Universidade Federal do Rio de Janeiro(UFRJ)Rio de JaneiroBrazil
  5. 5.Instituto Nacional de MetrologiaNormalização e Qualidade Industrial—InmetroRio de JaneiroBrazil
  6. 6.Centro de Ciências da Saúde (CCS)Universidade Federal do Rio de Janeiro (UFRJ)Rio de JaneiroBrazil

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