Skip to main content
Log in

A Study of Membrane Activity in Rat Prostate Cancer Cells: An Evaluation of the FM1-43 Dye Technique

  • Published:
Bioscience Reports

Abstract

A study was initiated to test whether the FM1-43 dye technique could beapplied to the study of endocytic membrane activity in two rodent prostatecancer (MAT-LyLu and AT-2) cell lines of markedly different metastaticability. The lipophilic dye FM1-43, which has frequently been used tomonitor endo/exocytic activity in excitable cells was employed. We found,as in excitable tissues, that both strongly metastatic (MAT-LyLu) andweakly metastatic (AT-2) cells in culture take up FM1-43 to give vesicularstaining of a variable pattern, which appeared to differ between the twocell lines. However, unlike excitable tissues, neither cell linesubsequently released the dye. Indeed, both cell lines retained the dyethrough several rounds of cell division suggesting that dye incorporatedby cells does not enter the endo/exocytotic cycle. Uptake of dye wasindependent of temperature, Na+/K+ gradients, pH or metabolism. Wesuggest that passive accumulation of FM1-43 can occur in cancer cells andshould not, automatically, be interpreted as evidence of endocytosis.

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

  • Aznavoorian, S., Murphy, A. N., Stetler-Stevenson, W. G., and Liotta, L. (1993) Molecular aspects of tumor cell invasion and metastasis. Cancer 71:1368–1383.

    PubMed  Google Scholar 

  • Betz, W. J., and Bewick, G. S. (1992) Optical analysis of synaptic vesicle recycling at the frog neuromuscular junction. Science 225:200–203.

    Google Scholar 

  • Betz, W. J., and Bewick, G. S. (1993) Optical monitoring of transmitter release and synaptic vesicle recycling at the neuromuscular junction. J. Physiology 460:287–309.

    Google Scholar 

  • Betz, W. J., Mao, F., and Bewick G. S. (1992) Activity-dependent and destaining of vertebrate motor nerve terminals. J. Neuroscience 12:363–375.

    Google Scholar 

  • Bowman, E. G., Siebers, A., and Altendorf, K. (1988) Bafilomycins: A class of inhibitors of membrane ATPases from microorganisms, animal cells and plant cells. Proc. Nat. Acad. Sci. USA 85:7972–7976.

    PubMed  Google Scholar 

  • Burgoyne, R. D., Morgan, A., and O'Sullivan, A. J. (1989) The control of cytoskeletal actin and exocytosis in intact and permeabilised adrenal chromaffin cells: Role of calcium and protein kinase. Cellular Signalling 1:323–344.

    PubMed  Google Scholar 

  • Foster, C. S., Cornford, P., Forsth, L., Djamgoz, M. B. A., and Ke, Y. (1999) The cellular and molecular basis of prostate cancer. Brit. J. Urology 83:171–194.

    Google Scholar 

  • Fraser, S. P., Salvador, V., and Djamgoz, M. B. A. (1998) Voltage-gated Na+ channel activity contributes to rodent prostate cancer cell migration in vitro. J. Physiology 513.P:131P.

    Google Scholar 

  • Fraser, S. P., Ding, Y., Liu, A., Foster, C. S., and Djamgoz, M. B. A. (1999) Tetrodotoxin suppresses morphological enhancement of the metastatic MAT-LyLu rat prostate cancer cell line. Cell and Tissue Research 295:505–512.

    PubMed  Google Scholar 

  • Gluck, S., Kelly, S., and Al-Awqati, Q. (1992) The proton translocating ATPases responsible for urinary acidification. J. Biological Chemistry 257:9230–9233.

    Google Scholar 

  • Grimes, J. A. et al. (1995) Differential expression of voltage-activated Na+ currents in two prostatic tumour cell lines: contribution to invasiveness in vitro. FEBS Letters 369:290–294.

    Google Scholar 

  • Grimes, J. A., and Djamgoz, M. B. A. (1998) Electrophysiological characterization of voltage-gated Na+ current expressed in the highly metastatic Mat-LyLu cell line of rat prostate cancer. J. Cellular Physiology 175:50–58.

    Google Scholar 

  • Houslay, M. D., and Stanley, K. K. (1982) Dynamics of Biological Membranes: Influence on Synthesis, Structure and Function. John Wiley and Sons, Chichester.

    Google Scholar 

  • Isaacs, J. T., Isaacs, W. B., Feitz, W. F. J., and Scheres, J. (1986) Establishment and characterization of seven Dunning rat prostatic cancer cell lines and their use in developing methods for predicting metastatic abilities of prostatic cancers. Prostate 9:261–281.

    PubMed  Google Scholar 

  • Katz, B. (1969) The Release of Neural Transmitter Substances. Liverpool Universit Press, Liverpool.

    Google Scholar 

  • Lagnado, L., Gomis, A., and Job, C. (1996) Continuous vesicle cycling in the synaptic terminal of retinal bipolar cells. Neuron 17:957–967.

    PubMed  Google Scholar 

  • Lagunoff, D., and Wan, H. (1974) Temperature dependence of mast cell histamine secretion. J. Cell Biology 61:809–811.

    Google Scholar 

  • Laniado, M. E. et al. (1997) Expression and functional analysis of Na+ channels in human prostate cancer cell lines and their contribution to invasiveness in vitro. Am. J. Path. 150:1213–1221.

    Google Scholar 

  • Meyer, T. and Hart, I. R. (1998) Mechanisms of tumour metastasis. European J. Cancer 34:214–221.

    Google Scholar 

  • Reuter, H. (1995) Measurement of exocytosis from single presynaptic nerve terminals reveal heterogeneous inhibition by Ca2+ channel blockers. Neuron 14:773–779.

    PubMed  Google Scholar 

  • Russell, P. J., Bennett, S., and Stricker, P. (1998) Growth factor involvement in progression of prostate cancer. Clinical Chemistry 44:705–723.

    PubMed  Google Scholar 

  • Ryan, T. A., Smith, S. J., and Reuter, H. (1996) The timing of synaptic vesicle endocytosis. Proc. Nat. Acad. Sci. USA 93:5567–5571.

    PubMed  Google Scholar 

  • Schacher, S., Holtzman, E., and Hood, D. C. (1976) Synaptic activity of frog retinal photoreceptors. A peroxidase uptake study. J. Cell Biology 70:178–192.

    Google Scholar 

  • Smith, P. H. et al. (1998) Sodium channel protein expression enhances the invasiveness of rat and human prostate cancer cells. FEBS Letters 423:19–24.

    Google Scholar 

  • Theodosis, D. T., Dreifuss, J. J., Harris, M. C., and Orci, L. (1976) Secretion related uptake of horseradish peroxidase in neurohypophysial axons. J. Cell Biology 70:294–303.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Mycielska, M.E., Schäfer, M., Fraser, S.P. et al. A Study of Membrane Activity in Rat Prostate Cancer Cells: An Evaluation of the FM1-43 Dye Technique. Biosci Rep 20, 31–40 (2000). https://doi.org/10.1023/A:1005531100458

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1005531100458

Navigation