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J. Biol. Chem., Vol. 278, Issue 14, 12222-12230, April 4, 2003
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From the Metabotropic glutamate receptors 5 (mGluR5)
are members of the growing group C G protein-coupled receptor
family. Widely expressed in mammalian brain, they are involved in
modulation of the glutamate transmission. By means of transfection of
mGluR5 receptors in COS-7 cells and primary hippocampal neurons in
culture followed by immunocytochemistry and quantitative image analysis
and by a biochemical assay, we have studied the internalization of
mGluR5 splice variants. mGluR5a and -5b were endocytosed in COS-7 cells as well as in axons and dendrites of cultured neurons. Endocytosis occurred even in the absence of receptor activity, because receptors mutated in the glutamate binding site were still internalized as well
as receptors in which endogenous activity had been inhibited by
an inverse agonist. We have measured a constitutive rate of endocytosis
of 11.7%/min for mGluR5a. We report for the first time the endocytosis
pathway of mGluR5. Internalization of mGluR5 is not mediated by
clathrin-coated pits. Indeed, inhibition of this pathway by Eps15
dominant negative mutants did not disturb their endocytosis. However,
the large GTPase dynamin 2 is implicated in the endocytosis of mGluR5
in COS-7. mGluR5 is the first shown member of the group C
G-protein coupled receptor family internalized by a nonconventional pathway.
The Metabotropic Glutamate Receptor mGluR5 Is Endocytosed by
a Clathrin-independent Pathway*
,,
"Physiologie Cellulaire de la Synapse,"
UMR 5091 CNRS/Université Bordeaux 2, Institut François
Magendie, Rue Camille Saint Saëns, 33077 Bordeaux Cedex, France,
§ "Mécanismes Moléculaires des Communications
Cellulaires," UPR CNRS 9023, CCIPE, 141 Rue de la Cardonille, 34094 Montpellier Cedex 5, France, and ¶ "Analyse d'Images
Quantitative," URA CNRS 1947, Institut Pasteur, 25 Rue du Dr. Roux,
75724 Paris Cedex, France
*
This work was supported by grants from the Conseil
Régional d'Aquitaine, the Association pour la Recherche
Médicale en Aquitaine.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed. Tel.:
33-5-57-57-40-89; Fax: 33-5-57-57-40-82; E-mail:
ahemar@u-bordeaux2.fr.
Copyright © 2003 by The American Society for Biochemistry and Molecular Biology, Inc.
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