Skip to main content
Log in

Programmed cell death: Cytochemical and X-ray microanalytical characterization of calcium compartments in neuromuscular junctions during the normal breakdown of the intersegmental muscles in the giant silkmothAntheraea polyphemus

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

Summary

Calcium stores were cytochemically demonstrated using a combined oxalate—pyroantimonate method in the neuromuscular junctions of the degenerating intersegmental muscles in the giant silkmothAntheraea polyphemus. The elemental composition of punctate precipitates of the reaction product was determined by electron probe X-ray microanalysis of unstained thin sections by energy-dispersive spectrometry and wavelength-dispersive spectrometry. The wavelength-dispersive spectra collected over terminal axons demonstrate a significant calcium signal and a trace of antimony.

During the rapid lytic phase of spontaneous muscle degeneration, the calcium punctate deposits were detected in presynaptic terminals in the following sites: the synaptic vesicles and the mitochondria. Calcium precipitates were also found in the dense bodies and the mitochondria encountered in the glial convolutions. No calcium deposit was seen in the synaptic clefts and intercellular spaces of the subsynaptic reticulum of type I and type II. A comparison of calcium to antimony ratios between the terminal axons and the sarcoplasmic lysosomes revealed highly significant differences (P<0.001). Such a variability of the calcium to antimony ratio may be related to different conditions of precipitation or antimony diffusion in the different cell compartments. It was concluded that such synaptic terminals do not appear damaged in spite of the muscle degeneration and presumably continue to perform vital functions while the muscles are no longer contractile 20 h after adult ecdysis.

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

  • BEAULATON, J. (1986) Programmed cell death. Cytochemical evidence for accumulation of calcium in mitochondria and its translocation into lysosomes: X-ray microanalysis in metamorphosing insect muscles.Histochem. J. 18, 527–36.

    Google Scholar 

  • BEAULATON, J. & LOCKSHIN, R. A. (1977) Ultrastructural study of the normal degeneration of the intersegmental muscles ofAntheraea polyphemus andManduca sexta (Insecta Lepidoptera) with particular reference to cellular autophagy.J. Morphol. 154, 39–58.

    Google Scholar 

  • BEAULATON, J. & LOCKSHIN, R. A. (1978) Programmed cell death. Ultrastructural study of neuromuscular relations during degeneration of the intersegmental muscles.Biol. Cell. 33, 169–74.

    Google Scholar 

  • BEAULATON, J., NICAISE, G., NICOLAS, G. & LOCKSHIN, R. A. (1986) Programmed cell death. X-ray microanalysis of calcium and zinc within the electron-dense droplets of the T system in insect muscles.Biol. Cell 56, 271–82.

    Google Scholar 

  • BLITZ, A. L., FINE, R. E. & TOSELLI, P. A. (1977) Evidence that coated vesicles isolated from brain are calcium-sequestering organelles resembling sarcoplasmic reticulum.J. Cell Biol. 75, 135–47.

    Google Scholar 

  • BORGERS, M. (1985) Morphological assessment of tissue protection. InCalcium Entry Blockers and Tissue Protection (edited by GODFRAIND, T.et al.) pp. 173–81. New York: Raven Press.

    Google Scholar 

  • BORGERS, M., DE BRABANDER, M., VAN REEMPTS, J., AWOUTERS, F. & JACOB, W. A. (1977) Intranuclear microtubules in lung mast cells of guinea pigs in anaphylactic shock.Lab. Invest. 37, 1–8.

    Google Scholar 

  • BORGERS, M. & PIPER, H. M. (1986) Calcium-shifts in anoxic cardiac myocytes. A cytochemical study.J. molec. cell. Cardiol. 18, 439–48.

    Google Scholar 

  • BORGERS, M., THONÉ, F. & NUETEN, J. M. (1981) The subcellular distribution of calcium and the effects of calcium-antagonists as evaluated with a combined oxalate—pyroantimonate technique.Acta Histochem. 24, 327–32.

    Google Scholar 

  • BORGERS, M., THONÉ, F., VERHEYEN, A. & TER KEURS, H. E. D. J. (1984) Localization of calcium in skeletal and cardiac muscle.Histochem. J. 16, 295–309.

    Google Scholar 

  • BORGERS, M., THONÉ, F., XHONNEUX, R. & FLAMENG, W. (1982) Shifts of calcium in the ischemic myocardium. A structural analysis. InProtection of Tissues against Hypoxia (edited by WAUQUIER, A.et al.), pp. 365–75. Amsterdam: Elsevier Biomedical Press.

    Google Scholar 

  • CAMPBELL, A. K. (1983)Intracellular Calcium: its Universal Role as Regulator. Chichester: John Wiley.

    Google Scholar 

  • CHANDLER, J. A. (1977)X-ray Microanalysis in the Electron Microscope. InPractical Methods in Electron Microscopy (edited by GLAUERT, A. M.) Vol. 5, Part II, pp. 317–547. Amsterdam: North-Holland.

    Google Scholar 

  • FARBER, J. L. (1981) The role of calcium in cell death.Life Sci. 29, 1289–95.

    Google Scholar 

  • HENKART, M. P., REESE, T. S. & BRINLEY, F. J. (1978) Endoplasmic reticulum sequesters calcium in the squid giant axon.Science 202, 1300–3.

    Google Scholar 

  • JENNINGS, R. B. & HAWKINS, H. K. (1980) Ultrastructural changes of acute myocardial ischemia. InDegradative Processes in Heart and Skeletal Muscle (edited by WILDENTHAL, K.) pp. 295–346. Amsterdam: Elsevier/North-Holland Biomedical Press.

    Google Scholar 

  • LAFON-CAZAL, M. (1978) Les neurotransmetteurs des Insectes.Ann. Biol. 17, 489–528.

    Google Scholar 

  • LOCKSHIN, R. A. (1973) Degeneration of insect intersegmental muscles: electrophysiological studies of populations of fibres.J. Insect Physiol. 19, 2359–72.

    Google Scholar 

  • LOCKSHIN, R. A. & BEAULATON, J. (1974) Programmed cell death. Cytochemical evidence for lysosomes during the normal breakdown of the intersegmental muscles.J. Ultrastruct. Res. 46, 43–62.

    Google Scholar 

  • LOCKSHIN, R. A. & BEAULATON, J. (1979) Programmed cell death. Electrophysiological and ultrastructural correlations in metamorphosing muscles of lepidopteran insects.Tissue & Cell 11, 803–19.

    Google Scholar 

  • MCGRAW, C. F., SOMLYO, A. P. & BLAUSTEIN, M. P. (1980) Localization of calcium in presynaptic nerve terminals. An ultrastructural and electron microprobe analysis.J. Cell Biol. 85, 228–41.

    Google Scholar 

  • MELDRUM, B. S., SIMON, R. P., SWAN, J., EVANS, M. C. & GRIFFITHS, T. (1985) Calcium loading of mitochondria in ischemia and status epilepticus: its reversibility and significance for pathological outcome. InCalcium Entry Blockers and Tissue Protection (edited by GODFRAIND, T.et al.), pp. 183–94. New York: Raven Press.

    Google Scholar 

  • OHARA, P. T., WADE, C. R. & LIEBERMAN, A. R. (1979) Calcium storage sites in axon terminals and other components of intact CNS tissue: studies with a modified pyroantimonate technique.J. Anat. 129, 869–70.

    Google Scholar 

  • SOMLYO, A. P., SOMLYO, A. V., SHUMAN, H., SCARPA, A., ENDO, M. & INESI, G. (1981) Mitochondria do not accumulate significant Ca concentrations in normal cells. InCalcium Phosphate Transport Across Biomembranes (edited by BRONNER, F. & PETERLIK, K.) pp. 87–93. New York: Academic Press.

    Google Scholar 

  • TRUMAN, J. W. & SCHWARTZ, L. M. (1982) Insect systems for the study of programmed neuronal death.Neurosci. Comment. 1, 66–72.

    Google Scholar 

  • TRUMP, B. F. & BEREZESKY, I. K. (1983) The role of calcium deregulation in cell injury and cell death.Surv. Synth. Pathol. Res. 2, 165–9.

    Google Scholar 

  • USHERWOOD, P. N. R. & CULL-CANDY, S. G. (1975) Pharmacology of somatic nerve—muscle synapses. InInsect Muscle (edited by USHERWOOD, P. N. R.), pp. 207–80. London: Academic Press.

    Google Scholar 

  • VAN REEMPTS, J. & BORGERS, M. (1982) Morphological assessment of pharmacological brain protection. InProtection of Tissues against Hypoxia (edited by WAUQUIER, A.), pp. 263–74. Amsterdam: Elsevier Biomedical Press.

    Google Scholar 

  • VAN REEMPTS, J., BORGERS, M. & OFFNER, F. (1982) Ultrastructural localization of calcium in the rat retina with a combined oxalate—pyroantimonate technique.Histochem. J. 14, 517–22.

    Google Scholar 

  • VOLMER, H. (1981) Calcium uptake by sarcoplasmic reticulum vesicles from the leg muscles ofBlaberus fuscus.Insect Biochem. 11, 233–8.

    Google Scholar 

  • WALZ, B. (1979) Subcellular calcium localization and ATP-dependent Ca2+—uptake by smooth endoplasmic reticulum in an invertebrate photoreceptor cell. An ultrastructural, cytochemical and X-ray microanalytical study.Eur. J. Cell Biol. 20, 83–91.

    Google Scholar 

  • WALZ, B. (1982) Calcium-sequestering smooth endoplasmic reticulum in retinula cells of the blowfly.J. Ultrastruct. Res. 81, 240–8.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Beaulaton, J. Programmed cell death: Cytochemical and X-ray microanalytical characterization of calcium compartments in neuromuscular junctions during the normal breakdown of the intersegmental muscles in the giant silkmothAntheraea polyphemus . Histochem J 20, 131–138 (1988). https://doi.org/10.1007/BF01746676

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01746676

Keywords

Navigation