Abstract
New quantitative insights on the native high order chromatin-DNA structure existing within interphase nuclei are obtained by monitoring the effects of two common well-characterized fixatives, glutaraldehyde and ethanol/acetic acid mixture, at the level of the intranuclear DNA distribution and structures. Reproducible distinct levels of DNA fluorescence intensity and their intranuclear distribution are apparent in unfixed and fixed thymocytes by using DAPI and quantitative optical microscopy based on a charge coupled device. The fluorescent histograms correlated with the calorimetric thermograms on the very same thymocytes fixed and unfixed, establish an unequivocal baseline for the different levels of structural organization of the chromatin within the intact nucleus; namely their number, DNA packing ratio and fiber diameter. A systematic comparison among all the numerous models, being so far proposed for the quinternary and quaternary levels of DNA folding, to identifies the rope or ribbon-like and the chromonema as the ones that best fit with the in situ distribution.
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References
Belmont AS, Braunfeld MB, Sedat JW & Agard DA (1989) Chromosoma (Berlin) 98: 129–143
Belmont AS, Sedat JW & Agard DA (1987) J. Cell Biol. 105: 77–92
Kendall FM, Beltrame F, Zietz S, Belmont AS & Nicolini C (1980) Cell Biophys. 2: 373–404
Kendall FM, Swenson R, Borun Y, Rowinski J & Nicolini C (1977) Science 196: 1106–1109
Manuelidis L & Chen TL (1990). Cytometry 11: 8–25
Nicolini C (1986) Biophysics and Cancer, Plenum Press, N.Y.
Nicolini C (1991) In Molecular Basis of Human Cancer. (pp. 73–100) Plenum Press, N.Y.
Belmont AS, Kendall FM, & Nicolini, C (1984) J. Cell Sci. 65: 123–138
Diaspro A, Bertolotto M, Vergani L & Nicolini C (1991) IEEE Transactions on Biomedical Engineering BME-38: 670–678
Nicolini C, Trefiletti V, Cavazza B, Cuniberti C, Patrone E, Carlo P & Brambilla G (1983) Science 219: 176–178
Nicolini C, & Kendall F (1977) Physiol. Chem. Phys. 9: 265–283
Nicolini C, Carlo P, Martelli A, Finollo R, Patrone E, Trefiletti V & Brambilla G (1982) J. Mol. Biol. 161: 155–175
Nicolini C, Cavazza B, Trefiletti V, Pioli F, Beltrame F, Brambilla G, Maraldi N, & Patrone E (1983) J. Cell Sci. 62: 103–115
Vergani L, Mascetti GC, Gavazzo P & Nicolini C (1992) Thermochimica Acta 206: 175–179
Nicolini C, Diaspro A, Bertolotto M, Facci P & Vergani L (1991) Biochemical and Biophysical Research Comm. 177: 1313–1318
Diaspro A, Sartore M, Nicolini C, (1990) Image Vision and Computing 8: 130–134
Diaspro A, Adami M, Nicolini C, (1990) Image Vision and Computing 8: 134–141
Agard DA, Hiraoka Y, Shaw P, Sedat JW, (1989) Cell Biol. 30: 353–377
Agard DA, Sedat JW (1980) Spie. 264: 110–117
Mascetti G, Vergani L, Diaspro A, Carrara S, Radicchi G & Nicolini C (1996) Cytometry 23: 110–119
Kendall FM, Beltrame F & Nicolini C (1979). In Chromatin Structure and Function pp. 265–292, N.Y.: Plenum Press
Dupraw E. (1965) Nature 206: 338–340
Dickerson RE, Drew HR, Conner B. Fratins A & Kopka ML (1982) Science 216: 475–478
Nicolini C (1983) Anticancer Research 3: 63–86
Mc Ghee JD & Felsenfeld G (1980) Annu. Rev. Biochem. 49: 1115–1128
Nicolini C, Facci P. & Alliata D (1996) Nanobiology, in press
Woodcock CL, Frado L & Rattner J (1984) J. Cell Biol. 99: 42–52
Richmond TJ, Finch JT, Rushton B, Rhodes D & Klug A(1984) Nature 311: 532–537
Finch T. & Klug A. (1976) Proc. Nat. Acad. Sci. USA 73: 1887–1889
Widom J & Klug A (1985) Cell 43: 207–213
Williams S, Athey BD, Muglia LJ, Schappe RS, Gough AH & Langmore JP (1986). Biophys. J. 49, 233–248
Marsden MPF & Laemmli UK (1979) Cell 17: 849–858
Adolph KW (1980) Exp. Cell Res. 125: 95–103
Nicolini C, Vernazza B, Chiabrera A, Maraldi IN, Capitani S (1984) J. Cell Sci. 72: 75–87
Rattner JB and Lin CC (1985) Cell 42: 291–296
Woodcock CL (1994) J. Cell Biol. 1: 11–19
Belmont AS & Bruce K (1994) J. Cell Biol. 127: 287–302
Giammasca PJ, Horowitz RA & Woodcock CL (1993) J. Cell Sci. 105: 551–561
Langmore JP & Paulson JR (1983) J. Cell Biol. 96: 1120–1131
Paulson JR & Laemmli UK (1977) Cell 12: 817
Sedat J & Manuelidis L (1978) Cold Spring Harbour Symp. Biol. 42: 331–345
Nagl W (1977) In: Rostl TL & Gifford EM (eds) Mechanism of cell division, Dowden, Hutchinson & Ross, Stroudsburg, Pa
Shaw P, Histochem. J. Deconvolution in 3Doptical microscopy 26 (1994) 687–694
Russo I, Barboro P, Alberti I, Parodi S, Balbi C, Allera C, Lazzarini G, Patrone E, Role of H1 in chromatin folding: a thermodinamical study of chromatin reconstitution by differential scanning calomrimetry, Biochemistry 34 (1995) 301–311
Balbi C, Abelmoschi M, Zunino A, Cuniberti C, Cavazza B, Barboro P and Patrone E (1988) The decondensation process of nuclear chromatin as investigated by Differential Scanning Calorimetry. Biochem. Pharmacol. 37: 1815
Touchetta NA, Cole RD (1985) Differential scanning calorimetry of nuclei reveals the loss of major structural features in chromatin by brief nuclease treatment, Proc. Natl. Acad. Sci. USA 82: 2642
Nicolini C, Trefiletti V, Cavazza B, Cuniberti C, Patrone E, Carlo P, Brambilla G (1983) Science 219: 176
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Nicolini, C., Carrara, S. & Mascetti, G. High order DNA structure as inferred by optical fluorimetry and scanning calorimetry. Mol Biol Rep 24, 235–246 (1997). https://doi.org/10.1023/A:1006861801216
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DOI: https://doi.org/10.1023/A:1006861801216