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. 1987 Jul;84(14):4910–4913. doi: 10.1073/pnas.84.14.4910

Hindered diffusion of inert tracer particles in the cytoplasm of mouse 3T3 cells.

K Luby-Phelps, P E Castle, D L Taylor, F Lanni
PMCID: PMC305216  PMID: 3474634

Abstract

Using fluorescence recovery after photobleaching, we have studied the diffusion of fluorescein-labeled, size-fractionated Ficoll in the cytoplasmic space of living Swiss 3T3 cells as a probe of the physical chemical properties of cytoplasm. The results reported here corroborate and extend the results of earlier experiments with fluorescein-labeled, size-fractionated dextran: diffusion of nonbinding particles in cytoplasm is hindered in a size-dependent manner. Extrapolation of the data suggests that particles larger than 260 A in radius may be completely nondiffusible in the cytoplasmic space. In contrast, diffusion of Ficoll in protein solutions of concentration comparable to the range reported for cytoplasm is not hindered in a size-dependent manner. Although we cannot at present distinguish among several physical chemical models for the organization of cytoplasm, these results make it clear that cytoplasm possesses some sort of higher-order intermolecular interactions (structure) not found in simple aqueous protein solutions, even at high concentration. These results also suggest that, for native cytoplasmic particles whose smallest radial dimension approaches 260 A, size may be as important a determinant of cytoplasmic diffusibility as binding specificity. This would include most endosomes, polyribosomes, and the larger multienzyme complexes.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Anderson J. L., Quinn J. A. Restricted transport in small pores. A model for steric exclusion and hindered particle motion. Biophys J. 1974 Feb;14(2):130–150. doi: 10.1016/S0006-3495(74)70005-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Axelrod D., Koppel D. E., Schlessinger J., Elson E., Webb W. W. Mobility measurement by analysis of fluorescence photobleaching recovery kinetics. Biophys J. 1976 Sep;16(9):1055–1069. doi: 10.1016/S0006-3495(76)85755-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Clegg J. S. Properties and metabolism of the aqueous cytoplasm and its boundaries. Am J Physiol. 1984 Feb;246(2 Pt 2):R133–R151. doi: 10.1152/ajpregu.1984.246.2.R133. [DOI] [PubMed] [Google Scholar]
  4. Elson E. L., Reidler J. A. Analysis of cell surface interactions by measurements of lateral mobility. J Supramol Struct. 1979;12(4):481–489. doi: 10.1002/jss.400120408. [DOI] [PubMed] [Google Scholar]
  5. Fulton A. B. How crowded is the cytoplasm? Cell. 1982 Sep;30(2):345–347. doi: 10.1016/0092-8674(82)90231-8. [DOI] [PubMed] [Google Scholar]
  6. GROTTE G. Passage of dextran molecules across the blood-lymph barrier. Acta Chir Scand Suppl. 1956;211:1–84. [PubMed] [Google Scholar]
  7. Inman J. K. Thymus-independent antigens: the preparation of covalent, hapten-ficoll conjugates. J Immunol. 1975 Feb;114(2 Pt 1):704–709. [PubMed] [Google Scholar]
  8. Jacobson K., Wojcieszyn J. The translational mobility of substances within the cytoplasmic matrix. Proc Natl Acad Sci U S A. 1984 Nov;81(21):6747–6751. doi: 10.1073/pnas.81.21.6747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. LAUFFER M. A. Theory of diffusion in gels. Biophys J. 1961 Jan;1:205–213. doi: 10.1016/s0006-3495(61)86884-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Larm O., Lindberg B., Svensson S. Studies on the length of the side chains of the dextran elaborated by Leuconostoc mesenteroides NRRL B-512. Carbohydr Res. 1971 Nov;20(1):39–48. doi: 10.1016/s0008-6215(00)84947-2. [DOI] [PubMed] [Google Scholar]
  11. Laurent T. C. Determination of the structure of agarose gels by gel chromatography. Biochim Biophys Acta. 1967 Mar 22;136(2):199–205. doi: 10.1016/0304-4165(67)90064-5. [DOI] [PubMed] [Google Scholar]
  12. Laurent T. C., Granath K. A. Fractionation of dextran and Ficoll by chromatography on Sephadex G-200. Biochim Biophys Acta. 1967 Mar 22;136(2):191–198. doi: 10.1016/0304-4165(67)90063-3. [DOI] [PubMed] [Google Scholar]
  13. Lepock J. R., Cheng K. H., Campbell S. D., Kruuv J. Rotational diffusion of TEMPONE in the cytoplasm of Chinese hamster lung cells. Biophys J. 1983 Dec;44(3):405–412. doi: 10.1016/S0006-3495(83)84314-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Luby-Phelps K., Lanni F., Taylor D. L. Behavior of a fluorescent analogue of calmodulin in living 3T3 cells. J Cell Biol. 1985 Oct;101(4):1245–1256. doi: 10.1083/jcb.101.4.1245. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Luby-Phelps K., Taylor D. L., Lanni F. Probing the structure of cytoplasm. J Cell Biol. 1986 Jun;102(6):2015–2022. doi: 10.1083/jcb.102.6.2015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mastro A. M., Babich M. A., Taylor W. D., Keith A. D. Diffusion of a small molecule in the cytoplasm of mammalian cells. Proc Natl Acad Sci U S A. 1984 Jun;81(11):3414–3418. doi: 10.1073/pnas.81.11.3414. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. OGSTON A. G., WOODS E. F. Molecular configuration of dextrans in aqueous solution. Nature. 1953 Jan 31;171(4344):221–222. doi: 10.1038/171221a0. [DOI] [PubMed] [Google Scholar]
  18. Peters R., Peters J., Tews K. H., Bähr W. A microfluorimetric study of translational diffusion in erythrocyte membranes. Biochim Biophys Acta. 1974 Nov 15;367(3):282–294. doi: 10.1016/0005-2736(74)90085-6. [DOI] [PubMed] [Google Scholar]
  19. Pollard T. D. Cytoskeletal functions of cytoplasmic contractile proteins. J Supramol Struct. 1976;5(3):317–334. doi: 10.1002/jss.400050306. [DOI] [PubMed] [Google Scholar]
  20. Porter K. R. The cytomatrix: a short history of its study. J Cell Biol. 1984 Jul;99(1 Pt 2):3s–12s. doi: 10.1083/jcb.99.1.3s. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. RENKIN E. M. Filtration, diffusion, and molecular sieving through porous cellulose membranes. J Gen Physiol. 1954 Nov 20;38(2):225–243. [PMC free article] [PubMed] [Google Scholar]
  22. Stossel T. P. The structure of cortical cytoplasm. Philos Trans R Soc Lond B Biol Sci. 1982 Nov 4;299(1095):275–289. doi: 10.1098/rstb.1982.0132. [DOI] [PubMed] [Google Scholar]
  23. Taylor D. L., Condeelis J. S. Cytoplasmic structure and contractility in amoeboid cells. Int Rev Cytol. 1979;56:57–144. doi: 10.1016/s0074-7696(08)61821-5. [DOI] [PubMed] [Google Scholar]
  24. Taylor D. L., Fechheimer M. Cytoplasmic structure and contractility: the solation--contraction coupling hypothesis. Philos Trans R Soc Lond B Biol Sci. 1982 Nov 4;299(1095):185–197. doi: 10.1098/rstb.1982.0125. [DOI] [PubMed] [Google Scholar]
  25. Wang Y. L., Lanni F., McNeil P. L., Ware B. R., Taylor D. L. Mobility of cytoplasmic and membrane-associated actin in living cells. Proc Natl Acad Sci U S A. 1982 Aug;79(15):4660–4664. doi: 10.1073/pnas.79.15.4660. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Wojcieszyn J. W., Schlegel R. A., Wu E. S., Jacobson K. A. Diffusion of injected macromolecules within the cytoplasm of living cells. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4407–4410. doi: 10.1073/pnas.78.7.4407. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Yguerabide J., Schmidt J. A., Yguerabide E. E. Lateral mobility in membranes as detected by fluorescence recovery after photobleaching. Biophys J. 1982 Oct;40(1):69–75. doi: 10.1016/S0006-3495(82)84459-7. [DOI] [PMC free article] [PubMed] [Google Scholar]

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