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. 1986 Jun 1;102(6):2015–2022. doi: 10.1083/jcb.102.6.2015

Probing the structure of cytoplasm

PMCID: PMC2114258  PMID: 2423529

Abstract

We have used size-fractionated, fluorescent dextrans to probe the structure of the cytoplasmic ground substance of living Swiss 3T3 cells by fluorescence recovery after photobleaching and video image processing. The data indicate that the cytoplasm of living cells has a fluid phase viscosity four times greater than water and contains structural barriers that restrict free diffusion of dissolved macromolecules in a size-dependent manner. Assuming these structural barriers comprise a filamentous meshwork, the combined fluorescence recovery after photobleaching and imaging data suggest that the average pore size of the meshwork is in the range of 300 to 400 A, but may be as small as 200 A in some cytoplasmic domains.

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Selected References

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  1. Blikstad I., Markey F., Carlsson L., Persson T., Lindberg U. Selective assay of monomeric and filamentous actin in cell extracts, using inhibition of deoxyribonuclease I. Cell. 1978 Nov;15(3):935–943. doi: 10.1016/0092-8674(78)90277-5. [DOI] [PubMed] [Google Scholar]
  2. Bray D., Thomas C. The actin content of fibroblasts. Biochem J. 1975 May;147(2):221–228. doi: 10.1042/bj1470221. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. 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]
  5. GROTTE G. Passage of dextran molecules across the blood-lymph barrier. Acta Chir Scand Suppl. 1956;211:1–84. [PubMed] [Google Scholar]
  6. Gershon N. D., Porter K. R., Trus B. L. The cytoplasmic matrix: its volume and surface area and the diffusion of molecules through it. Proc Natl Acad Sci U S A. 1985 Aug;82(15):5030–5034. doi: 10.1073/pnas.82.15.5030. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Heuser J. E., Kirschner M. W. Filament organization revealed in platinum replicas of freeze-dried cytoskeletons. J Cell Biol. 1980 Jul;86(1):212–234. doi: 10.1083/jcb.86.1.212. [DOI] [PMC free article] [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. Kim H., Binder L. I., Rosenbaum J. L. The periodic association of MAP2 with brain microtubules in vitro. J Cell Biol. 1979 Feb;80(2):266–276. doi: 10.1083/jcb.80.2.266. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Koppel D. E. Association dynamics and lateral transport in biological membranes. J Supramol Struct Cell Biochem. 1981;17(1):61–67. doi: 10.1002/jsscb.380170107. [DOI] [PubMed] [Google Scholar]
  11. Kreis T. E., Geiger B., Schlessinger J. Mobility of microinjected rhodamine actin within living chicken gizzard cells determined by fluorescence photobleaching recovery. Cell. 1982 Jul;29(3):835–845. doi: 10.1016/0092-8674(82)90445-7. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. LAURENT T. C., BJOERK I., PIETRUSZKIEWICZ A., PERSSON H. ON THE INTERACTION BETWEEN POLYSACCHARIDES AND OTHER MACROMOLECULES. II. THE TRANSPORT OF GLOBULAR PARTICLES THROUGH HYALURONIC ACID SOLUTIONS. Biochim Biophys Acta. 1963 Oct 29;78:351–359. doi: 10.1016/0006-3002(63)91645-7. [DOI] [PubMed] [Google Scholar]
  14. Lanni F., Ware B. R. Detection and characterization of actin monomers, oligomers, and filaments in solution by measurement of fluorescence photobleaching recovery. Biophys J. 1984 Jul;46(1):97–110. doi: 10.1016/S0006-3495(84)84002-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. 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]
  17. Lieska N., Yang H. Y., Goldman R. D. Purification of the 300K intermediate filament-associated protein and its in vitro recombination with intermediate filaments. J Cell Biol. 1985 Sep;101(3):802–813. doi: 10.1083/jcb.101.3.802. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. 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]
  20. 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]
  21. Penman S., Fulton A., Capco D., Ben Ze'ev A., Wittelsberger S., Tse C. F. Cytoplasmic and nuclear architecture in cells and tissue: form, functions, and mode of assembly. Cold Spring Harb Symp Quant Biol. 1982;46(Pt 2):1013–1028. doi: 10.1101/sqb.1982.046.01.094. [DOI] [PubMed] [Google Scholar]
  22. Peters R. Nucleo-cytoplasmic flux and intracellular mobility in single hepatocytes measured by fluorescence microphotolysis. EMBO J. 1984 Aug;3(8):1831–1836. doi: 10.1002/j.1460-2075.1984.tb02055.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Phillies G. D. Diffusion of bovine serum albumin in a neutral polymer solution. Biopolymers. 1985 Feb;24(2):379–386. doi: 10.1002/bip.360240206. [DOI] [PubMed] [Google Scholar]
  24. Pollard T. D. Molecular architecture of the cytoplasmic matrix. Kroc Found Ser. 1984;16:75–86. [PubMed] [Google Scholar]
  25. Pollard T. D., Selden S. C., Maupin P. Interaction of actin filaments with microtubules. J Cell Biol. 1984 Jul;99(1 Pt 2):33s–37s. doi: 10.1083/jcb.99.1.33s. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. 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]
  27. Ris H. The cytoplasmic filament system in critical point-dried whole mounts and plastic-embedded sections. J Cell Biol. 1985 May;100(5):1474–1487. doi: 10.1083/jcb.100.5.1474. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sato M., Wong T. Z., Brown D. T., Allen R. D. Rheological properties of living cytoplasm: a preliminary investigation of squid axoplasm (Loligo pealei). Cell Motil. 1984;4(1):7–23. doi: 10.1002/cm.970040103. [DOI] [PubMed] [Google Scholar]
  29. Schliwa M., van Blerkom J. Structural interaction of cytoskeletal components. J Cell Biol. 1981 Jul;90(1):222–235. doi: 10.1083/jcb.90.1.222. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Sims P. J., Waggoner A. S., Wang C. H., Hoffman J. F. Studies on the mechanism by which cyanine dyes measure membrane potential in red blood cells and phosphatidylcholine vesicles. Biochemistry. 1974 Jul 30;13(16):3315–3330. doi: 10.1021/bi00713a022. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. 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]
  33. 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]
  34. Valberg P. A., Albertini D. F. Cytoplasmic motions, rheology, and structure probed by a novel magnetic particle method. J Cell Biol. 1985 Jul;101(1):130–140. doi: 10.1083/jcb.101.1.130. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. 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]
  36. Wang Y. L., Taylor D. L. Preparation and characterization of a new molecular cytochemical probe: 5-iodoacetamidofluorescein-labeled actin. J Histochem Cytochem. 1980 Nov;28(11):1198–1206. doi: 10.1177/28.11.6107318. [DOI] [PubMed] [Google Scholar]
  37. Wolosewick J. J., Porter K. R. Microtrabecular lattice of the cytoplasmic ground substance. Artifact or reality. J Cell Biol. 1979 Jul;82(1):114–139. doi: 10.1083/jcb.82.1.114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. 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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