Abstract
To assess if membrane diffusion could affect the kinetics of receptor recruitment at adhesive contacts, we transfected neurons with green fluorescent protein-tagged immunoglobin cell adhesion molecules of varying length (25-180 kD), and measured the lateral mobility of single quantum dots bound to those receptors at the cell surface. The diffusion coefficient varied within a physiological range (0.1-0.5 microm(2)/s), and was inversely proportional to the size of the receptor. We then triggered adhesive contact formation by placing anti-green fluorescent protein-coated microspheres on growth cones using optical tweezers, and measured surface receptor recruitment around microspheres by time-lapse fluorescence imaging. The accumulation rate was rather insensitive to the type of receptor, suggesting that the long-range membrane diffusion of immunoglobin cell adhesion molecules is not a limiting step in the initiation of neuronal contacts.
MeSH Terms
Animals
Biophysics/methods
Cell Adhesion
Cell Adhesion Molecules/chemistry
Cell Membrane/metabolism
Cells, Cultured
Diffusion
Green Fluorescent Proteins/chemistry,metabolism
Hippocampus/metabolism
Immunoglobulins/chemistry
Kinetics
Microscopy, Fluorescence
Molecular Weight
Neurons/metabolism
Protein Binding
Rats
Time Factors
Transfection
Chemicals
Cell Adhesion Molecules
Immunoglobulins
NrCAM protein, rat
Green Fluorescent Proteins
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Thoumine Olivier
CNRS 5091, Université Bordeaux 2, Bordeaux, France. olivier.thoumine@pcs.u-bordeaux2.fr
Saint-Michel Edouard
Dequidt Caroline
Falk Julien
Rudge Rachel
Galli Thierry
Faivre-Sarrailh Catherine
Choquet Daniel
References (8)
8 references, click to expand
-
Immunoglobulin superfamily receptors: cis-interactions, intracellular adapters and alternative splicing regulate adhesion.
Curr Opin Cell Biol. 2001 Oct;13(5):611-8
PMID: 11544031
-
Ankyrin binding mediates L1CAM interactions with static components of the cytoskeleton and inhibits retrograde movement of L1CAM on the cell surface.
J Cell Biol. 2003 Aug 18;162(4):719-30
PMID: 12925712
-
NrCAM coupling to the cytoskeleton depends on multiple protein domains and partitioning into lipid rafts.
Mol Biol Cell. 2004 Oct;15(10):4695-709
PMID: 15254265
-
Regulation of N-cadherin dynamics at neuronal contacts by ligand binding and cytoskeletal coupling.
Mol Biol Cell. 2006 Feb;17(2):862-75
PMID: 16319177
-
Truncation mutants define and locate cytoplasmic barriers to lateral mobility of membrane glycoproteins.
Proc Natl Acad Sci U S A. 1994 Apr 12;91(8):3378-82
PMID: 8159755
-
Tyrosine phosphorylation at a site highly conserved in the L1 family of cell adhesion molecules abolishes ankyrin binding and increases lateral mobility of neurofascin.
J Cell Biol. 1997 May 5;137(3):703-14
PMID: 9151675
-
Structural mosaicism on the submicron scale in the plasma membrane.
Biophys J. 1998 Jan;74(1):297-308
PMID: 9449330
-
The lateral mobility of some membrane proteins is determined by their ectodomains.
Biophys J. 1992 Apr;62(1):92-4
PMID: 1318107