Skip to main content
Log in

Immunolocalization of ubiquitin in degenerating insect flight muscle

  • Papers
  • Published:
The Histochemical Journal Aims and scope Submit manuscript

Summary

Ubiquitin was localized by immunofluorescence microscopy during post-mating histolysis of fibrillar flight muscle in female fire ants, Solenopsis spp. Normal muscles, as well as histolysing muscles from artificially inseminated and haemolymph-injected females contained ubiquitin in association with nuclei, Z-lines, myofilaments and mitochondria. However, the density of the ubiquitin immunoreaction was markedly increased in the nuclei, Z-lines and mitochondria of degenerating tissues 6, 12 and 24 h post treatment. At these times the heaviest immunoreactivity for ubiquitin was seen in association with the nuclei, Z-lines and mitochondria. Immuno-controls, incubated in the absence of the primary antibody, showed no similar immunostaining. When insemination was preceded by the injection of actinomycin D, muscle degradation was significantly depressed after a 24-h period. Also, ubiquitin immunofluorescence was markedly reduced in tissues pre-treated with actinomycin D. These observations suggest that insemination increases the ubiquitination of specific myofibrillar proteins destined for degradation.

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

Access this article

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

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Ball, E., Karlik, C. C., Beall, C. J., Saville, D. L., Sparrow, J. C., Bullard, B. & Fyrberg, E. A. (1987) Arthrin, a myofibrillar protein of flight muscle, is an actin-ubiquitin conjugate Cell 51, 221–8.

    Google Scholar 

  • Ciechanover, A., Finely, D. & Varshavsky, A. (1984) The ubiquitin-dependent pathway and mechanisms of energy-dependent intracellular protein degradation. J. Cell Biochem. 24, 27–53.

    Google Scholar 

  • Davis, W. L., Jones, R. G. & Farmer, G. R. (1989) Insect hemolymph factor promotes histolysis in Solenopsis. Anat. Rec. 224, 473–8.

    Google Scholar 

  • Davis, W. L., Jacoby, B. H., Jones, R. G. & Goodman, D. B. P. (1993a) Superoxide formation preceding flight muscle degeneration in Solenopsis: fine structural cytochemistry and biochemistry. Histochem. J. 25, 478–490.

    Google Scholar 

  • Davis, W. L., Hagler, H. K., Jacoby, B. H., Konrad, R. J. & Goodman, D. B. P. (1993b) TNF-a induced changes in the fine structure, ion distribution and oxyradical formation in insect flight muscle. Histochem. J. (submitted for publication).

  • Goldstein, G., Scheid, M., Hammerling, U., Boyse, E. A., Schelsinger, D. H. & Niall, H. D. (1975) Isolation of a polypeptide that has lymphocyte differentiating properties and is probably present universally in living cells. Proc. Natl Acad. Sci. USA 75, 11–15.

    Google Scholar 

  • Haas, A. L. & Bright, P. M. (1985) The immunochemical detection and quantitation of intracellular ubiquitin-protein conjugates. J. Biol Chem. 260, 12 464–73.

    Google Scholar 

  • Herschko, A., Eytan, E., Ciechanover, A. & Haas, A. L. (1982) Immunochemical analysis of the turnover of the ubiquitin-protein conjugates in intact cells: relationship to the breakdown of abnormal proteins. J. Biol. Chem. 257, 13 964–70.

    Google Scholar 

  • Herschko, A. & Ciechanover, A. (1983) Mechanisms of intracellular protein breakdown. Ann. Rev. Biochem. 51, 335–64.

    Google Scholar 

  • Hilenski, L. L., Terracio, L., Haas, A. L. & Borg, T. K. (1992) Immunolocalization of ubiquitin conjugates at Z-bands and intercalated discs of rat cardiomyocytes in vitro and in vivo. J. Histochem. Cytochem. 40, 1037–42.

    Google Scholar 

  • Jones, R. G., Davis, W. L., Hung, A. C. F. & Vinson, S. B. (1978) Insemination-induced histolysis of the flight muscle in fire ants (Solenopsis, spp.): an ultrastructural study. Am. J. Anat. 151, 603–10.

    Google Scholar 

  • Jones, R. G., Davis, W. L. & Vinson, S. B. (1982) A histochemical and microanalysis study of calcium changes in insect flight muscle degeneration in Solenopsis, the queen fire ant. J. Histochem. Cytochem. 30, 293–304.

    Google Scholar 

  • Jones, R. G. & Davis, W. L. (1985a) Muscle membranes from early degeneration muscle cell fibers in Solenopsis are leaky to lanthanum: electron microscopy and x-ray analysis. Anat. Rec. 212, 123–8.

    Google Scholar 

  • Jones, R. G. & Davis, W. L. (1985b) Leupeptin, a protease inhibitor blocks insemination-induced flight muscle histolysis in the fire ant Solenopsis. Tissue Cell 17, 111–6.

    Google Scholar 

  • Knowles, S. E. & Ballard, F. J. (1976) Selective control of the degradation of normal and aberrant proteins in Reuber H35 hepatoma cells. Biochem. J. 156, 609–17.

    Google Scholar 

  • Libby, P. & Goldberg, A. L. (1978) Leupeptin, a protease inhibitor, decreases protein degradation in normal and diseased muscles. Science 199, 534–6.

    Google Scholar 

  • Mortimore, G. E. (1982) Mechanisms of cellular protein catabolism. Nutr. Rev. 40, 1–12.

    Google Scholar 

  • Obinata, T. (1981) Dynamics of protein turnover in vertebrate skeletal muscle. Muscle Nerve 4, 456–88.

    Google Scholar 

  • Pringle, J. W. S. (1949) The excitation and contraction of the flight muscles of insects. J. Physiol. 108, 225–32.

    Google Scholar 

  • Pringle, J. W. S. (1978) Stretch activation of muscle: function and mechanism. Proc. Roy. Soc. Lond. Ser. B 201, 107–31.

    Google Scholar 

  • Rechsteiner, M. (1988) Ubiquitin. New York: Plenum Press.

    Google Scholar 

  • Riley, D. A., Bain, J. L. W. & Haas, A. L. (1986) Increased ubiquitin conjugation of proteins during skeletal muscle atrophy. J. Cell Biol. 103, 401A.

    Google Scholar 

  • Riley, D. A., Bain, J. L. W., Ellis, S. & Haas, A. L. (1988) Quantitation and immunocytochemical localization of ubiquitin conjugates within rat red and white skeletal muscles. J. Histochem. Cytochem. 36, 621–32.

    Google Scholar 

  • Riley, D. A., Ilyina-Kakueva, E. L., Ellis, S., Bain, J. L.W., Slocum, G. R. & Sedlak, F. R. (1990) Skeletal muscle fibers, nerve, and blood vessel breakdown in space-flown rats. FASEB J. 4, 84–91.

    Google Scholar 

  • Sanger, J. W., Mittal, B. & Sanger, J. M. (1984) Analysis of myofibrillar structure and assembly using fluorescein labeled contractile proteins. J. Cell Biol. 98, 825–833.

    Google Scholar 

  • Schwartz, L. M. & Truman, J. W. (1982) Peptide and steroid regulation of muscle degeneration in an insect. Science 215, 1420–1.

    Google Scholar 

  • Schwartz, L. M. & Truman, J. W. (1983) Hormonal control rates of metamorphic development in the Tobacco Hookworm Manduca sexta. Dev. Biol. 99, 103–14.

    Google Scholar 

  • Schwartz, L. M., Kosz, L. & Kay, B. K. (1990) Gene activation is required for developmentally programmed cell death. Proc. Natl Acad. Sci. USA 87, 6594–8.

    Google Scholar 

  • Seufert, W. & Jentsch, S. (1990) Ubiquitin-conjugating enzymes UBC4 and UBC5 mediate selective degradation of short-lived and abnormal proteins. EMBO J. 9, 543–550.

    Google Scholar 

  • Stracher, A., Mcgowan, E. B. & Shafiq, S. A. (1978) Muscular dystrophy: inhibition of degeneration in vivo with protease inhibitors. Science 200, 50–51.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Davis, W.L., Jacoby, B.H. & Goodman, D.B.P. Immunolocalization of ubiquitin in degenerating insect flight muscle. Histochem J 26, 298–305 (1994). https://doi.org/10.1007/BF00157762

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00157762

Keywords

Navigation