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PMID:15596489

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Citation

Waharte, F, Brown, CM, Coscoy, S, Coudrier, E and Amblard, F (2005) A two-photon FRAP analysis of the cytoskeleton dynamics in the microvilli of intestinal cells. Biophys. J. 88:1467-78

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 microm3. 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 microm2 s(-1) with a mobile fraction of 60% for actin, and 4 microm2 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.

Links

PubMed PMC1305148 Online version:10.1529/biophysj.104.049619

Keywords

Actins/physiology; Actins/ultrastructure; Caco-2 Cells; Calmodulin-Binding Proteins/physiology; Calmodulin-Binding Proteins/ultrastructure; Cell Movement/physiology; Computer Simulation; Cytoskeleton/physiology; Cytoskeleton/ultrastructure; Enterocytes/physiology; Enterocytes/ultrastructure; Fluorescence Recovery After Photobleaching/methods; Humans; Microscopy, Fluorescence, Multiphoton/methods; Microvilli/physiology; Microvilli/ultrastructure; Models, Biological; Molecular Motor Proteins/physiology; Molecular Motor Proteins/ultrastructure; Myosin Heavy Chains; Myosin Type I

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

CHICK:MYO1A

located_in

GO:0016328: lateral plasma membrane

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

CHICK:MYO1A

located_in

GO:0005884: actin filament

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

CHICK:MYO1A

located_in

GO:0005902: microvillus

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

CHICK:MYO1A

enables

GO:0000146: microfilament motor activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

CHICK:MYO1A

part_of

GO:0005884: actin filament

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

CHICK:MYO1A

part_of

GO:0005902: microvillus

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

CHICK:MYO1A

part_of

GO:0016328: lateral plasma membrane

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

CHICK:MYO1A

enables

GO:0016887: ATPase activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

CHICK:MYO1A

involved_in

GO:0030048: actin filament-based movement

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete


See also

References

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