Summary
A microspectrofluorometric approach in conjunction with microelectrophoretic injection of glucose-6-P (G6P), was used to study the effect of cultivation in a benzo(a)pyrene (BP)-containing medium on the metabolization of BP, by EL2 ascites cells.
The fluorescence increase which follows the microinjection of G6P to single living cells. was studied in EL2 cells grown for 24-hours, 48-hours, 10 days and 57 days in a BP-containing medium, as well as EL2 cells grown for 16 days in presence of BP and placed then for 24 or 48 hours in a BP-free medium. The effects on the induction of Aryl Hydrocarbon Hydroxylase were observed for cells so treated, and it seems that for the latter the requirement of intracellular O2 for the metabolization of BP, has been verified. Furthermore, another important factor has emerged: the intracellular BP concentration. The various spectra are discussed in function of these considerations.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Alfred, L. J., Gelboin, H. V.: Benzpyrene hydroxylase induction by polycyclic hydrocarbons in hamster embryonic cells grown in vitro. Science 157, 75–76 (1967)
Gelboin, H. V., Kinoshita, N., Wiebel, F. J.: Microsomal hydroxylases: induction and role in polycyclic hydrocarbon carcinogenesis and toxicity. Fed. Prod. 31, 1298–1309 (1972)
Hansen, A. R., Fouts, J. R.: Some problems in Michaelis-Menten kinetic analysis of Benzypyrene Hydroxylase in hepatic microsomes from polycyclic hydrocarbon-pretreated animals. Chem.-Biol. Interactions 5, 167–182 (1972)
Kodama, Y., Bock, F. G.: Benzo(a)pyrene metabolizing enzyme activity of livers of various strains of mice. Cancer Res. 30, 1846–1849 (1970)
Kohen, E., Bengtsson, G., Salmon, J. M., Kohen, C.: Critical parameters in the response of single EL2 ascites cells to intracellular addition of glucose-6-P. A microspectrofluorometric analysis. (in prep.), (1974d)
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)
Kohen, E., Kohen, C., Thorell, B., Wagner, G.: Analysis of metabolic transients in intact cells by rapid microfluorometry. Mikrochimica Acta (Wien) 1972, 103 (1972a)
Kohen, E., Kohen, C., Thorell, B.: Rapid multichannel microfluorometry for metabolic and spectral studies in single living cells. Exp. Cell Res. 81, 477–481 (1973a)
Kohen, E., Kohen, C., Thorell, B., Salmon, J. M.: 32-msec scan of the NAD(P)H fluorescence spectrum in single living cells. Rev. sci. Instrum. 44, 1784–1785 (1973b)
Kohen, E., Thorell, B., Kohen, C., Salmon, J. M.: Microfluorometric monitoring of dehydrogenase systems in the intact cell. Techniques of Biochemical and Biophysical Morphology, vol. 2, pp. 157–195 (D. Glick & Rosenbaum, eds.). New York: Wiley, 1974a)
Kohen, E., Salmon, J. M., Kohen, C., Bengtsson, G., Thorell, B.: Rapid microspectrofluorometry for biochemical and metabolic studies in single living cells. BBA, in press (1974b)
Kohen, E., Salmon, J. M., Kohen, C., Bengtsson, G.: Microspectrofluorometric evaluation of the oxygen probe 1-pyrene butyric acid in single living cells. (in prep.), (1974c)
Kouri, E. R., Lubet, R. A., Brown, D. Q.: Quantitation of Aryl Hydrocarbon Hydroxylaes activity in individual fetal hamster cells in vitro. J. nat. Cancer Inst. 49, 993–1004 (1972)
Salmon, J. M., Kohen, E., Kohen, C., Bengtsson, G.: A microspectrofluorometric approach for the study of benzo(a)pyrene and dibenzo(ah)anthracene metabolisation in single living cells. Histochemistry, in press(1974a)
Salmon, J. M., Kohen, E., Kohen, C., Bengtsson, G.: The effect of intracellular oxygen on the metabolization of benzo(a)pyrene and benzo(k)fluoroanthene. A microspectrofluorometric analysis in single living cells. Histochemistry, in press (1974b)
Tarbell, D. S., Brooker, E. G., Seifert, P., Vanterpool, A., Claus, O., Conway, W.: Studies on the metabolic products obtained from mouse skin after painting with 3,4-Benzpyrene. Cancer Res. 16, 37–47 (1956)
Watanabe, M., Potter, R., van, Morris, H. P.: Benzene Hydroxylase activity and induction by methyl-cholanthrene in Morris hepatomas, in host livers, in adult liver and in rat liver during development. Cancer Res. 30, 263–273 (1970)
Wattenberg, L. W., Leong, J. L.: Inhibition of the carcinogenic action of benzopyrene by flavones. Cancer Res. 30, 1922–1925 (1970)
Weigert, F., Mottram, J. C.: The biochemistry of Benzpyrene. I. A survey and new methods of analysis. Cancer Res. 6, 97–109 (1946)
Winer, A. D., Schwert, G. W., Millar, D. B. S.: VI. Fluorometric measurements of the complex of enzymes and reduced diphosphopyridine nucleotides. J. Biol. Chem. 234, 1149–1154 (1959)
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Salmon, JM., Kohen, E., Kohen, C. et al. Microspectrofluorometric study of Benzo(a)pyrene metabolization in Benzo(a)pyrene-grown single living cells. Histochemistry 42, 85–98 (1974). https://doi.org/10.1007/BF00498481
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00498481