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Methods Mol Biol. 2009;464:13-26. doi: 10.1007/978-1-60327-461-6_2.

Mechanical properties of interphase nuclei probed by cellular strain application.

Author information

1
Department of Medicine, Cardiovascular Division, Brigham and Women's Hospital/Harvard Medical School, Cambridge, MA, USA.

Abstract

The mechanical properties of the interphase nucleus have important implications for cellular function and can reflect changes in nuclear envelope structure and/or chromatin organization. Mutations in the nuclear envelope proteins lamin A and C cause several human diseases, such as Emery-Dreifuss muscular dystrophy, and dramatic changes in nuclear stiffness have been reported in cells from lamin A/C-deficient mice. We have developed a cellular strain technique to measure nuclear stiffness in intact, adherent cells and have applied this experimental method to fibroblasts from mouse models of Emery-Dreifuss muscular dystrophy and to skin fibroblasts from laminopathy patients and healthy control subjects. The experimental protocol is based on measuring induced nuclear deformations in cells plated on a flexible silicone substrate; the nuclear stiffness can subsequently be inferred from the ratio of induced nuclear strain to the applied membrane strain. These experiments reveal that lamins A and C are important determinants of nuclear stiffness and that lamin mutations associated with muscular dystrophies and other laminopathies often result in disturbed nuclear stiffness that could contribute to the tissue-specific disease phenotypes.

PMID:
18951177
PMCID:
PMC4153730
DOI:
10.1007/978-1-60327-461-6_2
[Indexed for MEDLINE]
Free PMC Article

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