Skip to main content
SpringerLink
Log in

Gene expression patterns in the black blowfly (Phormia regina) as revealed by two-dimensional electrophoresis of proteins. I. Developmental stage-specific and sex-specific differences

  • Published:
Biochemical Genetics Aims and scope Submit manuscript

Abstract

The black blowfly,Phormia regina, has been implicated in human myiasis and as a contact vector of viral and bacterial diseases present in carrion to which female flies are attracted for egg deposition. Inbred strains ofP. regina are an excellent model system for studying gene expression in the developmental stages of such holometabolous dipteran parasites. However, information regarding gene and protein expression patterns inP. regina is limited. We used ISO-DALT high-resolution, two-dimensional electrophoresis with silver staining to establish fundamental protein maps for examination of the stage-specific gene expression patterns in the 615 most abundant proteins of the eggs, first- and third-instar larvae, pupae, and male and female adults. We also used a differential extraction technique to identify the major cuticular proteins of the adults. The results show 48 clearly identifiable stage-specific and sex-specific proteins. Thus, approximately 8% of the most abundant proteins exhibit developmental changes. These analyses serve as an initial data base for further studies of ontogenetic regulation, organellar origin, and physiologic function of the stage-specific proteins in the life cycle of these opportunistically parasitic dipterans.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Alexis, J. B., and Mittleman, R. E. (1988). An unusual case ofPhormia regina myiasis of the scalp.Am. J. Clin. Pathol. 90734.

    PubMed  CAS  Google Scholar 

  • Ali-Khan, F. E., and Ali-Khan, Z. (1975). A case of traumatic dermal myiasis in Quebec caused byPhormia regina (Meigen) (Diptera: Calliphoridae).Can. J. Zool. 531470.

    Article  Google Scholar 

  • Anderson, N. G., and Anderson, N. L. (1982). The human protein index.Clin. Chem. 28739.

    PubMed  CAS  Google Scholar 

  • Anderson, N. L., and Anderson, N. G. (1977). High resolution two-dimensional electrophoresis of human plasma proteins.Proc. Natl. Acad. Sci. USA 745421.

    PubMed  CAS  Google Scholar 

  • Anderson, N. L., and Anderson, N. G. (1984). Some perspectives on two-dimensional protein mapping.Clin. Chem. 301898.

    PubMed  CAS  Google Scholar 

  • Anderson, N. L., and Hickman, B. J. (1979). Analytical techniques for cell fractions. XXIV. Isoelectric point standards for two-dimensional electrophoresis.Anal. Biochem. 93312.

    PubMed  CAS  Google Scholar 

  • Anderson, N. L., Giometti, C. S., Gemmell, M. A., and Macy, M. A. (1984a). Protein variants in cells: Enumeration by protein indexing.Ann. N.Y. Acad. Sci. 428134.

    PubMed  CAS  Google Scholar 

  • Anderson, N. L., Hofmann, J. P., Gemmell, M. A., and Taylor, J. (1984b). Global approaches to quantitative analysis of gene expression patterns observed by use of two-dimensional electrophoresis.Clin. Chem. 281807.

    Google Scholar 

  • Anderson, N. L., Tracy, R. P., and Anderson, N. G. (1984c). High resolution two-dimensional electrophoretic mapping of plasma proteins. In Putnam, F. (ed.),The Plasma Proteins 2nd ed., Academic Press, New York, pp. 221–270.

    Google Scholar 

  • Bauer, A. C., and Levenbook, L. (1969). Fructose diphosphate aldolase during growth and development of the blowflyPhormia regina (Meigen).Comp. Biochem. Physiol. 28619.

    Article  PubMed  CAS  Google Scholar 

  • Beaver, P. C., Jung, R. C., and Cupp, E. W. (1984). Filth flies and myiasis-producing flies. InClinical Parasitology 9th ed., Lea & Febiger, Philadelphia, p. 693.

    Google Scholar 

  • Bedian, V., Summers, M. C., and Kauffman, S. A. (1988). Changes in protein synthetic activity in earlyDrosophila embryos mutant for the segmentation geneKruppel.Dev. Genet. 9699.

    Article  PubMed  CAS  Google Scholar 

  • Chen, P. S., and Levenbook, L. (1966). Studies on the haemolymph proteins of the blowflyPhormia regina. I. Changes in ontogenetic patterns.J. Insect Physiol. 121595.

    Article  PubMed  CAS  Google Scholar 

  • Chen, P. S., Kubli, E., and Hanimann, F. (1968). Auftrennung der freien ninhydrin-positiven stoffe inPhormia undDrosophila mittels zweidimensionaler hochspannung-selektrophorese.Rev. Suisse Zool. 75509.

    PubMed  CAS  Google Scholar 

  • Clegg, J. S., and Evans, D. R. (1961). The physiology of blood trehalase and its function during flight in the blowfly.J. Exp. Biol. 38771.

    CAS  Google Scholar 

  • Coulthart, M. B., and Singh, R. S. (1988a). Low genic variation in male-reproductive-tract proteins ofDrosophila melanogaster andD. simulans.Mol. Biol. Evol. 5167.

    PubMed  CAS  Google Scholar 

  • Coulthart, M. B., and Singh, R. S. (1988b). Differing amounts of genetic polymorphism in testes and male accessory glands ofDrosophila melanogaster andDrosophila simulans.Biochem. Genet. 26153.

    Article  PubMed  CAS  Google Scholar 

  • Daufeldt, J. A., and Harrison, H. H. (1984). Quality control and technical outcome of high resolution two-dimensional electrophoresis in a clinical laboratory setting.Clin. Chem. 301972.

    PubMed  CAS  Google Scholar 

  • Fleming, J. E., Melnikoff, P. S., and Bensch, K. G. (1984). Identification of mitochondrial proteins on two-dimensional electrophoresis gels of extracts of adultDrosophila melanogaster.Biochim. Biophys. Acta 802340.

    CAS  Google Scholar 

  • Fleming, J. E., Quattrocki, E., Latter, G., Miguel, J., Marcuson, R., Zuckerkandl, E., and Bensch, K. G. (1986). Age-dependent changes in proteins ofDrosophila melanogaster.Science 2311157.

    PubMed  CAS  Google Scholar 

  • Friedman, S. (1960a). The purification and properties of trehalase isolated fromPhormia regina Meigen.Arch. Biochem. Biophys. 87252.

    Article  PubMed  CAS  Google Scholar 

  • Friedman, S. (1960b). Occurrence of trehalase-6-phosphatase inPhormia regina Meigen.Arch. Biochem. Biophys. 88339.

    Article  PubMed  CAS  Google Scholar 

  • Friedman, S. (1961). Inhibition of trehalase activity in the hemolymph ofPhormia regina.Arch. Biochem. Biophys. 93550.

    Article  PubMed  CAS  Google Scholar 

  • Gemmell, M. A., and Anderson, N. L. (1982). Lymphocyte, monocyte, and granulocyte proteins compared by use of two-dimensional electrophoresis.Clin. Chem. 271062.

    Google Scholar 

  • Giometti, C. S., and Anderson, N. L. (1984). Tropomyosin heterogeneity in human cells.J. Biol. Chem. 25914113.

    PubMed  CAS  Google Scholar 

  • Giometti, C. S., Danon, M. J., and Anderson, N. G. (1983). Human muscle proteins: Analysis by two-dimensional electrophoresis.Neurology 331152.

    PubMed  CAS  Google Scholar 

  • Giometti, C. S., Gemmell, M. A., Nance, S. L., Tollaksen, S. L., and Taylor, J. (1987). Detection of heritable mutations in protein expression.J. Biol. Chem. 26212764.

    PubMed  CAS  Google Scholar 

  • Goldenthal, M. J., McKenna, K. A., and Joslyn, D. J. (1991). The mitochondrial DNA (mtDNA) of the blowflyPhormia regina: Restriction map and gene localization.Biochem. Genet. 291.

    Article  PubMed  CAS  Google Scholar 

  • Green, L. A. D., Kaplan, M. P., and Liem, R. K. H. (1991). Kinesin heavy chain from bovine brain andDrosophila appear to be highly homologous molecules.J. Neurosci. Res. 28151.

    Article  PubMed  CAS  Google Scholar 

  • Guevara, J., Johnston, D. A., Ramagli, L. S., Martin, B. A., Capetillo, S., and Rodriguez, L. V. (1982). Quantitative aspects of silver deposition in proteins resolved in complex polyacrylamide gels.Electrophoresis 3197.

    Article  CAS  Google Scholar 

  • Gutzeit, H. O., and Gehring, W. J. (1979). Localized synthesis of specific proteins during oogenesis and early embryogenesis inDrosophila melanogaster.Wilhelm Roux Arch. 187151.

    Article  Google Scholar 

  • Hall, R. D., Anderson, P. C., and Clark, D. P. (1986). A case of human myiasis caused byPhormia regina.J. Med. Entomol. 23578.

    PubMed  CAS  Google Scholar 

  • Harrison, H. H. (1984). Improved record keeping and photography of silver-stained two-dimensional electrophoretograms by way of “XRD images.”Clin. Chem. 301981.

    PubMed  CAS  Google Scholar 

  • Hudson, A. (1958). The effect of flight on the taste threshold and carbohydrate utilization ofPhormia regina Meigen.J. Insect Physiol. 1293.

    Article  CAS  Google Scholar 

  • Hurlimann, R., and Chen, P. S. (1974). Ontogenetische veranderungen des enzymmusters in der haemolymphe vonPhormia regina.Rev. Suisse Zool. 81648.

    PubMed  CAS  Google Scholar 

  • Imajoh, S. (1981). Application of two-dimensional electrophoresis to the analysis of speciation inAnopheles hyrcanus complex.Seikagaku (Biochemistry) 53159.

    CAS  Google Scholar 

  • Kim, B. K. (1988). Phylogenetic relationships of the seven species of theDrosophila auraria complex by two-dimensional electrophoresis.Korean J. Genet. 1077.

    Google Scholar 

  • Kingan, T. G., and Hildebrand, J. G. (1989). Sexually dimorphic polypeptides in developing antennal sensory neurons of an insect.J. Neurosci. 91951.

    PubMed  CAS  Google Scholar 

  • Kominz, D. R., Maruyama, K., Levenbook, L., and Lewis, M. (1962). Tropomyosin, myosin, and actin from the blowfly,Phormia regina.Biochim. Biophys. Acta 63106.

    Article  PubMed  CAS  Google Scholar 

  • Lee, C. Y., Charles, D., Bronson, D., Griffin, M., and Bennett, L. (1979). Analysis of mouse andDrosophila proteins by two-dimensional gel electrophoresis.Mol. Gen. Genet. 176303.

    Article  PubMed  CAS  Google Scholar 

  • Lee, T. J., and Park, J. W. (1990). Genetic relationships among the five species of the subgenusHirtodrosophila genusDrosophila.Genetica 1271.

    Google Scholar 

  • Levenbook, L., Bauer, A. C., and Shigematsu, H. (1973). Fructose diphosphate aldolase during growth and development of the blowfly,Phormia regina (Meigen). III. Physicochemical comparison of the larval and adult muscle enzymes.Arch. Biochem. Biophys. 157615.

    Article  Google Scholar 

  • MacConkey, E. H., Taylor, B. J., and Phan, D. (1979). Human heterozygosity: A new estimate.Proc. Natl. Acad. Sci. USA 767951.

    Google Scholar 

  • Marmaras, V. J., and Tsakas, S. (1988). Temporally regulated protein synthesis in cultured haemocytes of the Mediterranean fruit flyCeratitis capitata during larval and prepupal development: Internalization of larval serum proteins into the haemocytes.Dev. Biol. 129294.

    Article  PubMed  CAS  Google Scholar 

  • Matthews, K. A., Miller, D. F. B., and Kaufman, T. C. (1989). Developmental distribution of RNA and protein products of theDrosophila α-tubulin gene family.Dev. Biol. 13245.

    Article  PubMed  CAS  Google Scholar 

  • Miller, S. G., and Huettel, M. D. (1986). Mitochondrial biogenesis during spermatogenesis inHeliothis virescens, H. sublflexa, and their male-sterile backcross hybrids.Arch. Insect Biochem. Physiol. 3363.

    Article  CAS  Google Scholar 

  • Mogami, K., Fujita, S. C., and Hotta, Y. (1982). Identification ofDrosophila indirect flight muscle myofibrillar proteins by means of two-dimensional electrophoresis.J. Biochem. 91643.

    PubMed  CAS  Google Scholar 

  • Nash, W. G., Kumerow, H. N., and Merril, C. R. (1983). Identification of aYellow gene-specific protein inDrosophila melanogaster by two-dimensional electrophoresis.Biochem. Genet. 211135.

    Article  PubMed  CAS  Google Scholar 

  • Neel, J. V., Baier, L., and Hanash, S. M. (1985). Frequency of polymorphisms for alleles encoding for liver proteins of domesticated mice.J. Hered. 76314.

    PubMed  CAS  Google Scholar 

  • O'Farrell, P. H. (1975). High resolution two-dimensional electrophoresis of proteins.J. Biol. Chem. 2504007.

    PubMed  Google Scholar 

  • Ramagli, L. S., Womack, J. E., and Rodriguez, L. V. (1990). Genetic analysis of nonhistone chromosomal protein inheritance in recombinant inbred mouse strains using two-dimensional electrophoresis.Biochem. Genet. 28123.

    Article  PubMed  CAS  Google Scholar 

  • Ramesh, S. R., and Kalisch, W. E. (1988). Taxonomic identification ofDrosophila nasuta subgroup strains by glue protein analysis.Genetica 7863.

    Article  PubMed  Google Scholar 

  • Rosenblum, B. B., Neel, J. V., Hanash, S. M., Joseph, J. L., and Yew, N. (1984). Identification of genetic variants in erythrocyte lysate by two-dimensional electrophoresis.Am. J. Hum. Genet. 36601.

    PubMed  CAS  Google Scholar 

  • Sakoyama, Y., and Okubo, S. (1981). Two-dimensional gel patterns of protein species during development ofDrosophila embryos.Dev. Biol. 81361.

    Article  PubMed  CAS  Google Scholar 

  • Santaren, J. F. (1990). Towards establishing a protein database ofDrosophila.Electrophoresis 11254.

    Article  PubMed  CAS  Google Scholar 

  • Santaren, J. F., and Garcia-Bellido, A. (1990). High-resolution two-dimensional gel analysis of proteins in wing imaginal discs: A database ofDrosophila.Exp. Cell Res. 189169.

    Article  PubMed  CAS  Google Scholar 

  • Seaquist, E. R., Henry, T. R., Cheong, E., and Theologides, D. (1983).Phormia regina myiasis in a malignant wound.Minn. Med. 66409.

    PubMed  CAS  Google Scholar 

  • Shymala, B. V., and Ranganath, H. A. (1990). Biochemical phylogeny of seven Indian species of theMontium subgroup ofDrosophila.Genetica 8171.

    Article  Google Scholar 

  • Silvert, D. J., Doctor, J., Quesada, L., and Fristrom, J. W. (1984). Pupal and larval cuticle proteins ofDrosophila melanogaster.Biochemistry 235767.

    Article  PubMed  CAS  Google Scholar 

  • Spicer, G. S. (1988). Molecular evolution among someDrosophila species groups as indicated by two-dimensional electrophoresis.J. Mol. Evol. 27250.

    Article  PubMed  CAS  Google Scholar 

  • Takai, K., and Kanda, T. (1986). Phylogenetic relationships among theAnopheles hyrcanus species group based on the degree of hybrid development and comparison with phylogenies by other methods.Jpn. J. Genet. 61295.

    Google Scholar 

  • Takana-Ohmuro, H., Hirose, G., and Mikawa, T. (1983). Separation and identification ofDrosophila myosin light chains.J. Biochem. 211135.

    Google Scholar 

  • Tanaka, Y., Maruyama, K., Mikawa, T., and Hotta, Y. (1988). Identification of calcium binding proteins in two-dimensional gel electrophoretic patterns ofDrosophila thorax and their distribution in two types of muscle.J. Biochem. 104489.

    PubMed  CAS  Google Scholar 

  • Thorsrud, A. K., Vatn, M. H., and Jellum, E. (1982). Two-dimensional electrophoretic patterns of proteins of normal mucosa, polyps, and carcinomas of the large intestine.Clin. Chem. 28884.

    PubMed  CAS  Google Scholar 

  • Tollaksen, S. L., Anderson, N. L., and Anderson, N. G. (1984).Operation of the ISO-DALT System 7th ed., U.S. Dept. Energy Publ. ANL-BIM-84-1, Argonne Natl. Lab., 9700 South Cass Avenue, Argonne, IL 60439.

    Google Scholar 

  • Trumbly, R. J., and Jarry, B. (1983). Stage-specific protein synthesis during early embryogenesis inDrosophila melanogaster.EMBO J. 21281.

    PubMed  CAS  Google Scholar 

  • Tsai, Y. C., Harrison, H. H., Lee, C., Daufeldt, J. A., Oliver, L., and Grayhack, J. T. (1984). Systematic characterization of human prostatic fluid proteins with two-dimensional electrophoresis.Clin. Chem. 302026.

    PubMed  CAS  Google Scholar 

  • Walton, K. E., Styer, D., and Gruenstein, E. I. (1984). Genetic polymorphism in normal human fibroblasts as analyzed by two-dimensional polyacrylamide electrophoresis.J. Biol. Chem. 2547951.

    Google Scholar 

  • Westerbrink, K., Havsteen, B., and Groenier, K. (1983). Numerical taxonomy of two-dimensional protein maps: A rational approach to tumor characterization. In Stathakos, D. (ed.),Electrophoresis ′82 Walter de Gruyter, Berlin, pp. 423–433.

    Google Scholar 

  • Willard, K. E., and Anderson, N. G. (1981). Two-dimensional analysis of human lymphocyte proteins. I. An assay for lymphocyte effectors.Clin. Chem. 271327.

    PubMed  CAS  Google Scholar 

  • Young, D. A. (1984). Advantages of separations on “giant” two-dimensional gels for detection of physiologically relevant changes in the expression of protein gene-products.Clin. Chem. 302104.

    PubMed  CAS  Google Scholar 

  • Zebe, E. (1958). Studies on glycolytic enzymes in insect muscle.Int. Congr. Entomol. Montreal 2371.

    Google Scholar 

  • Zuckerkandl, E. (1986). Quantitative protein profiling: Determining lexotypes.J. Theoret. Biol. 121185.

    CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Pathology, The University of Chicago Pritzker School of Medicine, 5841 South Maryland Avenue, Box 146, 60637, Chicago, Illinois

    Harold H. Harrison

  2. Department of Biology, Rutgers University at Camden, 08102, Camden, New Jersey

    Dennis J. Joslyn

Authors
  1. Harold H. Harrison
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Dennis J. Joslyn
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Financial support was provided by the Sprague Institute (H. Harrison), UC Medlabs TRSP Fund (H. Harrison), New Jersey State Mosquito Control Commission, New Jersey Department of Environmental Protection (D. Joslyn), and Rutgers University Research Council (D. Joslyn), and an equipment loan by Argonne National Laboratory (Contract No. W-31-109-ENG-38).

Rights and permissions

Reprints and permissions

About this article

Cite this article

Harrison, H.H., Joslyn, D.J. Gene expression patterns in the black blowfly (Phormia regina) as revealed by two-dimensional electrophoresis of proteins. I. Developmental stage-specific and sex-specific differences. Biochem Genet 29, 559–576 (1991). https://doi.org/10.1007/BF02426871

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02426871

Key words

Search

Navigation