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.
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.
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.
Anderson, N. G., and Anderson, N. L. (1982). The human protein index.Clin. Chem. 28739.
Anderson, N. L., and Anderson, N. G. (1977). High resolution two-dimensional electrophoresis of human plasma proteins.Proc. Natl. Acad. Sci. USA 745421.
Anderson, N. L., and Anderson, N. G. (1984). Some perspectives on two-dimensional protein mapping.Clin. Chem. 301898.
Anderson, N. L., and Hickman, B. J. (1979). Analytical techniques for cell fractions. XXIV. Isoelectric point standards for two-dimensional electrophoresis.Anal. Biochem. 93312.
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.
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.
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.
Bauer, A. C., and Levenbook, L. (1969). Fructose diphosphate aldolase during growth and development of the blowflyPhormia regina (Meigen).Comp. Biochem. Physiol. 28619.
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.
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.
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.
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.
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.
Coulthart, M. B., and Singh, R. S. (1988a). Low genic variation in male-reproductive-tract proteins ofDrosophila melanogaster andD. simulans.Mol. Biol. Evol. 5167.
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.
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.
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.
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.
Friedman, S. (1960a). The purification and properties of trehalase isolated fromPhormia regina Meigen.Arch. Biochem. Biophys. 87252.
Friedman, S. (1960b). Occurrence of trehalase-6-phosphatase inPhormia regina Meigen.Arch. Biochem. Biophys. 88339.
Friedman, S. (1961). Inhibition of trehalase activity in the hemolymph ofPhormia regina.Arch. Biochem. Biophys. 93550.
Gemmell, M. A., and Anderson, N. L. (1982). Lymphocyte, monocyte, and granulocyte proteins compared by use of two-dimensional electrophoresis.Clin. Chem. 271062.
Giometti, C. S., and Anderson, N. L. (1984). Tropomyosin heterogeneity in human cells.J. Biol. Chem. 25914113.
Giometti, C. S., Danon, M. J., and Anderson, N. G. (1983). Human muscle proteins: Analysis by two-dimensional electrophoresis.Neurology 331152.
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.
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.
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.
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.
Gutzeit, H. O., and Gehring, W. J. (1979). Localized synthesis of specific proteins during oogenesis and early embryogenesis inDrosophila melanogaster.Wilhelm Roux Arch. 187151.
Hall, R. D., Anderson, P. C., and Clark, D. P. (1986). A case of human myiasis caused byPhormia regina.J. Med. Entomol. 23578.
Harrison, H. H. (1984). Improved record keeping and photography of silver-stained two-dimensional electrophoretograms by way of “XRD images.”Clin. Chem. 301981.
Hudson, A. (1958). The effect of flight on the taste threshold and carbohydrate utilization ofPhormia regina Meigen.J. Insect Physiol. 1293.
Hurlimann, R., and Chen, P. S. (1974). Ontogenetische veranderungen des enzymmusters in der haemolymphe vonPhormia regina.Rev. Suisse Zool. 81648.
Imajoh, S. (1981). Application of two-dimensional electrophoresis to the analysis of speciation inAnopheles hyrcanus complex.Seikagaku (Biochemistry) 53159.
Kim, B. K. (1988). Phylogenetic relationships of the seven species of theDrosophila auraria complex by two-dimensional electrophoresis.Korean J. Genet. 1077.
Kingan, T. G., and Hildebrand, J. G. (1989). Sexually dimorphic polypeptides in developing antennal sensory neurons of an insect.J. Neurosci. 91951.
Kominz, D. R., Maruyama, K., Levenbook, L., and Lewis, M. (1962). Tropomyosin, myosin, and actin from the blowfly,Phormia regina.Biochim. Biophys. Acta 63106.
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.
Lee, T. J., and Park, J. W. (1990). Genetic relationships among the five species of the subgenusHirtodrosophila genusDrosophila.Genetica 1271.
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.
MacConkey, E. H., Taylor, B. J., and Phan, D. (1979). Human heterozygosity: A new estimate.Proc. Natl. Acad. Sci. USA 767951.
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.
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.
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.
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.
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.
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.
O'Farrell, P. H. (1975). High resolution two-dimensional electrophoresis of proteins.J. Biol. Chem. 2504007.
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.
Ramesh, S. R., and Kalisch, W. E. (1988). Taxonomic identification ofDrosophila nasuta subgroup strains by glue protein analysis.Genetica 7863.
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.
Sakoyama, Y., and Okubo, S. (1981). Two-dimensional gel patterns of protein species during development ofDrosophila embryos.Dev. Biol. 81361.
Santaren, J. F. (1990). Towards establishing a protein database ofDrosophila.Electrophoresis 11254.
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.
Seaquist, E. R., Henry, T. R., Cheong, E., and Theologides, D. (1983).Phormia regina myiasis in a malignant wound.Minn. Med. 66409.
Shymala, B. V., and Ranganath, H. A. (1990). Biochemical phylogeny of seven Indian species of theMontium subgroup ofDrosophila.Genetica 8171.
Silvert, D. J., Doctor, J., Quesada, L., and Fristrom, J. W. (1984). Pupal and larval cuticle proteins ofDrosophila melanogaster.Biochemistry 235767.
Spicer, G. S. (1988). Molecular evolution among someDrosophila species groups as indicated by two-dimensional electrophoresis.J. Mol. Evol. 27250.
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.
Takana-Ohmuro, H., Hirose, G., and Mikawa, T. (1983). Separation and identification ofDrosophila myosin light chains.J. Biochem. 211135.
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.
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.
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.
Trumbly, R. J., and Jarry, B. (1983). Stage-specific protein synthesis during early embryogenesis inDrosophila melanogaster.EMBO J. 21281.
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.
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.
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.
Willard, K. E., and Anderson, N. G. (1981). Two-dimensional analysis of human lymphocyte proteins. I. An assay for lymphocyte effectors.Clin. Chem. 271327.
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.
Zebe, E. (1958). Studies on glycolytic enzymes in insect muscle.Int. Congr. Entomol. Montreal 2371.
Zuckerkandl, E. (1986). Quantitative protein profiling: Determining lexotypes.J. Theoret. Biol. 121185.
Author information
Authors and Affiliations
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
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/PL00020698
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/PL00020698