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Time-Resolved Spectroscopy of mitochondria, cells and tissues under normal and pathological conditions

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Abstract

In this study, the detailed dependence of light scattering on tissue architecture and intracellular composition has been investigated. Firstly, we simulated the reduced scattering coefficient (μs′) of the rat liver using the Mie theory, the Rayleigh-Debye-Gans approximation and electron microscopy data. Then, the reduced scattering coefficient of isolated rat liver mitochondria, isolated hepatocytes and various rat tissues (i.e. perfused liver, brain, muscle, tumors) was measured at 780 nm by using time-resolved spectroscopy and a sample-substitution protocol. The comparison of the isolated mitochondria data with the isolated hepatocyte and whole liver measurements suggests that the mitochondrial compartment is the primary factor for light propagation in hepatic tissue, thus strengthening the relevance of the preliminary theoretical study. Nevertheless, the possibility that other intracellular components, such as peroxisomes and lysosomes, interfere with light propagation in rat liver is discussed. Finally, we demonstrate that light scattering in normal rat tissues and tumors is roughly proportional to the mitochondrial content, according to estimates of the mitochondrial protein content of the tissues.

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References

  1. Wilson BC, Sevick E, Patterson MS, Chance B: Time-dependent optical spectroscopy and imaging for biomedical applications. Proc IEEE 80: 918–930, 1992

    Google Scholar 

  2. Patterson MS, Chance B, Wilson BC: Time resolved reflectance and transmittance for the non-invasive measurement of tissue optical properties. Appl Opt 28: 2331–2336, 1989

    Google Scholar 

  3. Sevick EM, Chance B, Leigh J, Nioka S, Maris M: Quantitation of timeand frequency-resolved optical spectra for the determination of tissue oxygenation. Anal Biochem 195: 330–351, 1991

    Google Scholar 

  4. Chance B, Leigh J, Miyake H, Smith D, Nioka S, Greenfeld R, Finlander M, Kaufmann K, Levy W, Young M, Cohen P, Yoshioka H, Boretsky R: Comparison of time-resolved and un-resolved measurements of deoxyhemoglobin in brain. Proc Natl Acad Sci USA 85: 4971–4975, 1988

    Google Scholar 

  5. Chance B, Nioka S, Kent J, McCully K, Fountain M, Greenfeld R, Holtom G: Time-resolved spectroscopy of hemoglobin and myoglobin in resting and ischemic muscle. Anal Biochem 174: 698–707, 1988

    Google Scholar 

  6. Kerker M: The Scattering of Light, and other Electromagnetic Radiation. Academic Press, New York, USA, 1969, pp 1–666

    Google Scholar 

  7. Ishimaru A: Wave Propagation and Scattering in Random Media. Academic Press, New York, USA, 1978, pp 1–572

    Google Scholar 

  8. Kerker M: Elastic and inelastic light scattering in flow cytometry. Cytometry 4: 1–10, 1983

    Google Scholar 

  9. Steinkamp JA: Flow cytometry. Rev Sci Instrum 55: 1375–1400, 1984

    Google Scholar 

  10. Beauvoit B, Liu H, Kang K, Kaplan PD, Miwa M, Chance B: Characterization of absorption and scattering properties for various yeast strains by time-resolved spectroscopy. Cell Biophys 23: 1–16, 1993

    Google Scholar 

  11. Hoffmann HP, Avers C: Mitochondrion of yeast: Ultrastructural evidence for one giant, branched organelle per cell. Nature 181: 749–751, 1973

    Google Scholar 

  12. Miyakawa I, Aoi H, Sando N, Kuroiwa T: Fluorescence microscopic studies of mitochondrial nucleoids during meiosis and sporulation in the yeast, Saccharomyces cerevisiae. J Cell Sci 66: 21–38, 1984

    Google Scholar 

  13. Stevens B: Mitochondrial structure. In: JN Strathern, EW Jones, JR Broach (eds). The Molecular Biology of the Yeast Saccharomyces cerevisiae. Life Cycle and Inheritance. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, USA, 1981, pp 471–504

    Google Scholar 

  14. Else PL, Hulbert AJ: An allometric comparison of the mitochondria of mammalian and reptilian tissues: The implications for the evolution of endothermy. J Comp Physiol B 156: 3–11, 1985

    Google Scholar 

  15. Schwerzmann K, Hoppeler H, Kayar SR, Weibel ER: Oxidative capacity of muscle and mitochondria: Correlation of physiological, biochemical, and morphometric characteristics. Proc Natl Acad Sci USA 86: 1583–1587, 1989

    Google Scholar 

  16. Takahashi M, Hood DA: Chronic stimulation-induced changes in mitochondria and performance in rat skeletal muscle. J Appl Physiol 74: 934–941, 1993

    Google Scholar 

  17. Jakovcic S, Swift HH, Gross NJ, Rabinowitz M: Biochemical and stereological analysis of rat liver mitochondria in different thyroid states. J Cell Biol 77: 887–901, 1978

    Google Scholar 

  18. Pedersen PL: Tumor mitochondria and the bioenergetics of cancer cells. Prog Exp Tumor Res 22: 190–274, 1978

    Google Scholar 

  19. Chen LB: Fluorescent labeling of mitochondria. Meth Cell Biol 29: 103–123, 1989

    Google Scholar 

  20. Smiley ST, Reers M, Mottola-Hartshorn C, Lin M, Chen A, Smith TW, Steele GD, Chen LB: Intracellular heterogeneity in mitochondrial membrane potentials revealed by a J-aggregate-forming lipophilic cation JC-1. Proc Natl Acad Sci USA 88: 3671–3675, 1991

    Google Scholar 

  21. Lemasters JJ, Di Guiseppi J, Nieminen AL, Herman B: Blebbing, free Ca2+ and mitochondrial membrane potential preceding cell death in hepatocytes. Nature 325: 78–81, 1987

    Google Scholar 

  22. Reber BFX, Somogyi R, Stucki JW: Hormone-induced intracellular calcium oscillations and mitochondrial energy supply in single hepatocytes. Biochim Biophys Acta 1018: 190–193, 1990

    Google Scholar 

  23. Hess R, Staubli W, Riess W: Nature of the hepatomegalic effect produced by ethyl-chlorophenoxy-isobutyrate in the rat. Nature 27: 856–858, 1965

    Google Scholar 

  24. Tandler B, Hoppel CL: Studies on giant mitochondria. Ann NY Acad Sci 488: 65–81, 1986

    Google Scholar 

  25. Susuki K: Giant hepatic mitochondria: Production in mice fed with cuprizone. Science 163: 81–82, 1969

    Google Scholar 

  26. Liu H, Miwa M, Beauvoit B, Wang NG, Chance B: Characterization of absorption and scattering properties of small-volume biological samples using time-resolved spectroscopy. Anal Biochem 213: 378–385, 1993

    Google Scholar 

  27. Berry MN, Edwards AM, Barrit GJ, Grivell MB, Halls HJ, Gannon BJ, Friend DS: Isolated hepatocytes. Preparation, properties and applications. In: RH Burdon, PH Knippenberg (eds). Laboratory Techniques in Biochemistry and Molecular Biology. Elsevier, New York, USA, 1991, vol. 21, pp 1–460

    Google Scholar 

  28. Johnson D, Lardy H: Isolation of liver or kidney mitochondria. Meth Enzymol 10: 94–96, 1967

    Google Scholar 

  29. Clark JB, Nicklas WJ: The metabolism of rat brain mitochondria. Preparation and characterization. J Biol Chem 245: 4724–4731, 1970

    Google Scholar 

  30. Moreadith RW, Fiskum G: Isolation of mitochondria from ascites tumor cells permeabilized with digitonin. Anal Biochem 137: 360–367, 1984

    Google Scholar 

  31. Rodriguez-Vico F, Martinez-Cayuela M, Garcia-Peregrin E, Ramirez H: A procedure for eliminating interferences in the Lowry method of protein determination. Anal Biochem 183: 275–278, 1989

    Google Scholar 

  32. Hatefi Y, Stiggall DL: Preparation and properties of succinate:ubiquinone oxidoreductase (complex II). Meth Enzymol 53: 21–27, 1978

    Google Scholar 

  33. Ross WT, Cardell RR: Proliferation of smooth endoplasmic reticulum and induction of microsomal drug-metabolizing enzymes after ether or halothane. Anesthesiology 48: 325–331, 1978

    Google Scholar 

  34. Brown BR, Sagalyn AM: Hepatic microsomal enzyme induction by inhalation of anesthetics: Mechanism in the rat. Anesthesiology 40: 152–161, 1974

    Google Scholar 

  35. Berge RK, Skrede S, Farstad M: Effects of clofibrate on the intracellular localization of palmitoyl-CoA hydrolase and palmitoyl-L-carnitine hydrolase in rat liver. FEBS Lett 124: 43–47, 1981

    Google Scholar 

  36. Hardeman D, Zomer HWM, Schutgens RBH, Tager JM, van den Bosch H: Effect of peroxisome proliferation on ether phospholipid biosynthesizing enzymes in rat liver. Int J Biochem 22: 1413–1418, 1990

    Google Scholar 

  37. Graaff R, Aarnoudse JG, Zijp JR, Sloot PMA, de Mul FFM, Greve J, Koelink MH: Reduced light-scattering properties for mixtures of spherical particles: A simple approximation derived from Mie calculation. Appl Opt 31: 1370–1376, 1992

    Google Scholar 

  38. Reynolds L, Johnson C, Ishimaru A: Diffusive reflectance from a finite blood medium: To the modelling of fiber optic catheters. Appl Opt 15: 2059–2067, 1976

    Google Scholar 

  39. Steinke JM, Shepherd AP: Comparison of Mie theory and the light scattering of red blood cells. Appl Opt 27: 4027–4033, 1988

    Google Scholar 

  40. Loud AV: A quantitative stereological description of the ultrastructure of normal rat liver parenchymal cell. J Cell Biol 37: 27–39, 1968

    Google Scholar 

  41. Weibel ER, Staubli W, Gnagi HR, Hess FA: Correlated morphometric and biochemical studies on the liver cell. I. Morphometric model, stereologic methods, and normal morphometric data for rat liver. J Cell Biol 42: 68–84, 1969

    Google Scholar 

  42. Blouin A, Bolender RP, Weibel ER: Distribution of organelles and membranes between hepatocytes and non-hepatocytes in the rat liver parenchyma. A stereological study. J Cell Biol 72: 441–455, 1977

    Google Scholar 

  43. Bolin FP, Preuss LE, Taylor RC, Ference RJ: Refractive index of some mammalian tissues using a fiber optic cladding method. Appl Opt 28: 2297–2303, 1989

    Google Scholar 

  44. Cross DA, Latimer P: Angular dependence of scattering from Escherichia coli cells. Appl Opt 11: 1225–1228, 1972

    Google Scholar 

  45. Fujime S, Takasaki-Ohsita M, Miyamoto S: Dynamic light scattering from polydisperse suspensions of large spheres. Characterization of isolated secretory granules. Biophys J 54: 1179–1184, 1988

    Google Scholar 

  46. Sloot PMA, Hoekstra AG, Figdor CG: Osmotic response of lymphocytes measured by means of forward light scattering: Theoretical considerations. Cytometry 9: 636–641, 1988

    Google Scholar 

  47. Gear ARL, Bednarek JM: Direct counting and sizing of mitochondria in solution. J Cell Biol 54: 325–345, 1972

    Google Scholar 

  48. Schwerzmann K, Cruz-Orive LM, Eggman R, Sanger A, Weibel ER: Molecular architecture of the inner membrane of mitochondria from rat liver: A combined biochemical and stereological study. J Cell Biol 102: 97–103, 1986

    Google Scholar 

  49. Flatmark T, Kryvi H, Tangeras A: Induction of megamitochondria by cuprizone (biscyclohexanone oxaldihydrazone). Evidence for an inhibition of the mitochondrial division process. Eur J Cell Biol 23: 141–148, 1980

    Google Scholar 

  50. Wakabayashi T, Asano M, Kurono C, Ozawa T: Mechanism of the formation of megamitochondria induced by copper-chelating agents. II. Isolation and some properties of megamitochondria from the cuprizone-treated mouse liver. Acta Path Jap 25: 39–49, 1975

    Google Scholar 

  51. Kurup CKR, Aithal HN, Ramasarma T: Increase of hepatic mitochondria on administration of ethyl alpha-p-chlorophenoxyisobutyrate to the rat. Biochem J 116: 773–779, 1970

    Google Scholar 

  52. Anthony LE, Schmucker DL, Mooney JS, Jones AL: A quantitative analysis of fine structure and drug metabolism in livers of clofibratetreated young adult and retired breeder rats. J Lipids Res 19: 151–163, 1978

    Google Scholar 

  53. Lipsky NG, Pedersen PL: Perturbation by clofibrate of mitochondrial levels in animals cells. J Biol Chem 257: 1473–1481, 1982

    Google Scholar 

  54. Myers MW, Bosmann HB: Mitochondrial protein content and enzyme activity of Reuber hepatoma H35. Cancer Res 34: 1989–1994, 1974

    Google Scholar 

  55. Matsuno T: Pathway of glutamate oxidation and its regulation in the HuH13 line of human hepatoma cells. J Cell Physiol 148: 290–294, 1991

    Google Scholar 

  56. Loncar D, Afzelius BA, Cannon B: Epididymal white adipose tissue after cold stress in rats. I. Non-mitochondrial changes. J Ultrastr Mol Str Res 101: 109–122, 1988

    Google Scholar 

  57. Kitai T, Beauvoit B, Chance B: Optical determination of fatty change of the graft liver with near-infrared time-resolved spectroscopy. Transplantation 62: 642–647, 1996

    Google Scholar 

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Beauvoit, B., Chance, B. Time-Resolved Spectroscopy of mitochondria, cells and tissues under normal and pathological conditions. Mol Cell Biochem 184, 445–455 (1998). https://doi.org/10.1023/A:1006855716742

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