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© The Rockefeller University Press, 0021-9525/1997//1459 $5.00
The Journal of Cell Biology, Volume 137, Number 7, , 1997 1459-1468

Article

Chromatin Dynamics in Interphase Nuclei and Its Implications for Nuclear Structure



James R. Abney*,{ddagger}, Bryan Cutler*, Misty L. Fillbach§, Daniel Axelrod||, and Bethe A. Scalettar*

* Department of Physics, {ddagger} Northwestern School of Law, and § Department of Mathematics, Lewis & Clark College, Portland, Oregon 97219; and || Biophysics Research Division and Department of Physics, University of Michigan, Ann Arbor, Michigan 48109

Translational dynamics of chromatin in interphase nuclei of living Swiss 3T3 and HeLa cells was studied using fluorescence microscopy and fluorescence recovery after photobleaching. Chromatin was fluorescently labeled using dihydroethidium, a membrane-permeant derivative of ethidium bromide. After labeling, a laser was used to bleach small (~0.4 µm radius) spots in the heterochromatin and euchromatin of cells of both types. These spots were observed to persist for >1 h, implying that interphase chromatin is immobile over distance scales >=0.4 µm. Over very short times (<1 s), a partial fluorescence recovery within the spots was observed. This partial recovery is attributed to independent dye motion, based on comparison with results obtained using ethidium homodimer-1, which binds essentially irreversibly to nucleic acids. The immobility observed here is consistent with chromosome confinement to domains in interphase nuclei. This immobility may reflect motion-impeding steric interactions that arise in the highly concentrated nuclear milieu or outright attachment of the chromatin to underlying nuclear substructures, such as nucleoli, the nuclear lamina, or the nuclear matrix.

1. Abbreviation used in this paper: D, diffusion coefficient.

Please address all correspondence to Bethe A. Scalettar, Department of Physics, Lewis & Clark College, Portland, OR 97219. Tel.: (503) 768-7585. Fax: (503) 768-7369. e-mail: bethe{at}lclark.edu

James R. Abney's current address is Kolisch Hartwell Dickinson McCormack & Heuser, PC, Patent, Trademark and Copyright Attorneys, 520 S.W. Yamhill Street, Suite 200, Portland, OR 97204.


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