Advertisement

Histochemistry

, Volume 47, Issue 4, pp 291–302 | Cite as

A preliminary microspectrofluorometric study of NAD(P) reduction in diBenzo(a, e) Fluoranthene-treated single living cells

  • Jean-Marie Salmon
  • Elli Kohen
  • Cahide Kohen
  • P. Viallet
  • F. Zajdela
Article

Summary

The fluorescence increase, due to NAD(P) reduction, following microelectrophoretic injection of glucose 6-P (G6P) into EL2 and NCTC 8739 single living cells treated with diBenzo(ae)Fluoranthene (diB(ae)F) and non-treated, has been studied with a rapid microspectrofluorometer. This study shows the enhanced capacity of treated cells to utilize larger doses (6–10 times more) of G6P than control cells. The time course of the return to the initial fluorescence level is essentially related to the magnitude of the injection dose. There are alterations (e.g. red & blue shifts) in the fluorescence spectrum of diB(ae)F-treated cells before injection and in the increase spectrum after injection of G6P, as compared to the same spectra in the diB(ae)F-untreated cells. This is discussed in reference to the metabolization of diB(ae)F as an alternative pathway for the reoxidation of NAD(P)H.

Keywords

Public Health Glucose Living Cell Control Cell Fluorescence Spectrum 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Arcos, J.C., Argus, M.F.: Chemical induction of cancer. Structural bases and biological mechanisms, Vol. II A, p. 28. New York: Academic Press 1974Google Scholar
  2. Chance, B., Schoener, B.: Spectrophotometric and kinetic studies of flavoproteins in tissues, cell suspensions, mitochondria and their fragments. In: Flavins and flavoproteins, BBA Library (E.C. Slater, ed.), Vol. 15, pp. 496–528. Amsterdam: Elsevier Publishing Co. 1966Google Scholar
  3. Estabrook, R.W., Franklin, M., Baron, J., Shigematsu, A., Hildebrandt, A.: Properties of membrane bound electron transfer complex of the hepatic endoplasmic reticulum associated with drug metabolism. In: Drugs and cell regulation (E. Mihich, ed.), pp. 227–257. New York: Academic Press 1971Google Scholar
  4. Evans, V.J., Andresen, W.F.: Effect of serum on spontaneous neoplastic transformations in vitro. Nat. Cancer Inst. 37, 247–249 (1966)Google Scholar
  5. Evans, V.J., Bryant, J.C., Keer, H.A., Schilling, E.L.: Chemically defined media for cultivation of long term cell strains from four mammalian species. Exp. Cell Res. 36, 439–474 (1964)Google Scholar
  6. Galeotti, T., Van Rossum, G.D.V., Mayer, D.H., Chance, B.: On the fluorescence of NAD(P)H in whole-cell preparations of tumours and normal tissues. Europ. J. Biochem. 17, 485–496 (1970)Google Scholar
  7. Gelboin, H.V.: A microsome dependent binding of benzo (a) pyrene to DNA. Cancer Res. 29, 1272–1276 (1969)Google Scholar
  8. Gelboin, H.V., Kinoshita, N., Wiebel, F.: Microsomal hydroxylases: induction and role in polycyclic hydrocarbon carcinogenesis and toxicity. Fed. Proc. 31, 1298–1309 (1972)Google Scholar
  9. Kohen, E., Bengtsson, G., Salmon, J.M., Kohen, C.: The investigation of critical parameters in the glycolytic response of single living cells by rapid microspectrofluorometric analysis. Microchim. Acta (Wien) 249 (1976I)Google Scholar
  10. Kohen, E., Hirschberg, J.G., Kohen, C., Wouters, A.W., Pearson, A., Salmon, J.M., Thorell, B.: Multichannel microspectrofluorometry for topographic and spectral analysis of NAD(P)H fluorescence in single living cells. Biochim. biophys. Acta (Amst.) 396, 149–154 (1975a)Google Scholar
  11. Kohen, E., Kohen, C., Jenkins, W.: The influence of microelectrophoretically introduced metabolites on pyridine nucleotide reduction in giant tissue culture ascites cells. Exp. Cell Res. 44, 175–194 (1966)Google Scholar
  12. Kohen, E., Kohen, C., Thorell, B., Salmon, J.M.: 32 msec scan of NAD(P)H fluorescence spectrum in single living cells. Rev. Sci. Instrum. 44, 1784–1788 (1973)Google Scholar
  13. Kohen, E., Kohen, C., Thorell, B., Wagener, G.: Analysis of metabolic transients in intact cells by rapid microfluorometry. Mikrochim. Acta (Wien) 103–114 (1972)Google Scholar
  14. Kohen, E., Thorell, B., Kohen, C., Salmon, J.M.: Microfluorometric monitoring of dehydrogenase systems in the intact cells. Techniques of biochemical and biophysical morphology (D. Glick and R. Rosenbaum, eds.), Vol. 2, pp. 157–195. New York: John Wiley & Sons 1975b)Google Scholar
  15. Lu, A.Y.H., Levin, W., West, S., Jacobson, M., Ryan, D., Kuntzman, R., Conney, A.M.: The role of cytochrome P-450 and P-448 in drug and steroid hydroxylations. Ann N.Y. Acad. Sci. 212, 156–174 (1972)Google Scholar
  16. Mannering, G.J.: Role of substrate binding to P-450 hemoproteins in drug metabolism. In: Drugs and cell regulation (E. Mihich, ed.), pp. 197–225. New York: Academic Press 1971Google Scholar
  17. Nebert, D.W., Bausserman, L.L.: Aryl hydrocarbon hydroxylase induction in cell culture as a function of gene expression. Ann N.Y. Acad. Sci. 179, 561–579 (1971)Google Scholar
  18. Olson, G.G.: Applications of an optical multichannel analyzer. Amer. Lab. 4, 57–63 (1971)Google Scholar
  19. Orten, J.M., Neuhaus, O.W.: In: Biochemistry, 8th ed., pp. 196–213. St. Louis: C.V. Mosby Co. 1970Google Scholar
  20. Salmon, J.M., Kohen, E., Kohen, C., Bengtsson, G.: A microspectrofluorometric approach for the study of benzo (a) pyrene and dibenzo (a, h) anthracene metabolisation in single living cells. Histochemistry 42, 61–74 (1974a)Google Scholar
  21. Salmon, J.M., Kohen, E., Kohen, C., Bengtsson, G.: The effect of intracellular oxygen on the metabolisation of benzo (a) pyrene and benzo (k) fluoranthene. A microspectrofluorometric analysis in single living cells. Histochemistry 42, 75–84 (1974b)Google Scholar
  22. Salmon, J.M., Kohen, E., Kohen, C., Bengtsson, G.: Microspectrofluorometric study of benzo (a) pyrene metabolisation in benzo (a) pyrene-grown single living cells. Histochemistry 42, 85–98 (1974c)Google Scholar
  23. Sanford, K.K., Fox, C.H., Handleman, S.L., Jones, G.M.: XIth Int. Cancer Congress, 62. Florence, Italy, 1974Google Scholar
  24. Wainio, W.W.: In the mammalian mitochondrial respiratory chain, p. 152. New York: Academic Press 1970Google Scholar
  25. Winer, A.D., Schwert, G.W., Millar, D.B.S.: Lactic dehydrogenase. IV. Fluorimetric measurement of the complex of enzyme and reduced diphosphopyridine nucleotides. J. biol. Chem. 234, 1149–1154 (1959)Google Scholar

Copyright information

© Springer-Verlag 1976

Authors and Affiliations

  • Jean-Marie Salmon
    • 1
  • Elli Kohen
    • 2
    • 3
    • 4
  • Cahide Kohen
    • 2
    • 3
    • 4
  • P. Viallet
    • 1
  • F. Zajdela
    • 5
  1. 1.Laboratoire de Chimie Physique, CentreUniversitaire de PrepignanPerpignanFrance
  2. 2.Papanicolaou Cancer Research InstituteMiamiUSA
  3. 3.Clinical Faculty. Department of PathologySchool of Medicine University of MiamiMiamiUSA
  4. 4.Adjunct Faculty, Department of PhysiologySchool of Medicine University of MiamiMiamiUSA
  5. 5.Unité de Physiologie Cellulaire-U-22 Faculte des Sciences Institut National de la Santé et de la Recherche MedicaleOrsayFrance

Personalised recommendations