NDLI: A Two-Photon FRAP Analysis of the Cytoskeleton Dynamics in the Microvilli of Intestinal Cells

Content Provider	PubMed Central
Author	François, Waharte Brown, Claire M. Coscoy, Sylvie Coudrier, Evelyne Amblard, François
Copyright Year	2005
Abstract	The molecular structure of the brush-border of enterocytes has been investigated since the 1980s, but the dynamics of this highly specialized subcellular domain have been difficult to study due to its small size. To perform a detailed analysis of the dynamics of cytoskeleton proteins in this domain, we developed two-photon fluorescence recovery after photobleaching and a theoretical framework for data analysis. With this method, fast dynamics of proteins in the microvilli of the brush border of epithelial intestinal cells can be measured on the millisecond timescale in volumes smaller than 1 μm3. Two major proteins of the cytoskeleton of the microvilli, actin and myosin 1a (Myo1a; formerly named brush border myosin I), are mobile in the brush-border of Caco-2 cells, an enterocyte-like cellular model. However, the mobility of actin is very different from that of Myo1a and they appear to be unrelated (diffusion coefficient of 15 μm2 s−1 with a mobile fraction of 60% for actin, and 4 μm2 s−1 with a mobile fraction of 90% for Myo1a). Furthermore, we show for the first time, in vivo, that the dynamics of Myo1a in microvilli reflect its motor activity.
Related Links	http://dx.doi.org/10.1529/biophysj.104.049619
Ending Page	1478
Page Count	12
Starting Page	1467
File Format	PDF
ISSN	00063495
e-ISSN	15420086
Journal	Biophysical Journal
Issue Number	2
Volume Number	88
Language	English
Publisher	Biophysical Society
Publisher Date	2005-02-01
Access Restriction	Open
Rights Holder	Biophysical Society
Subject Keyword	Biophysics Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Biophysics

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