The Journal of Neuroscience, April 23, 2008, 28(17):4336-4349; doi:10.1523/JNEUROSCI.4379-07.2008
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Cellular/Molecular
Dynamics of Somatostatin Type 2A Receptor Cargoes in Living Hippocampal Neurons
Benjamin Lelouvier,1,2
Gianluca Tamagno,3,4
Angela M. Kaindl,1,2
Alexandre Roland,5
Vincent Lelievre,1,2
Virginia Le Verche,1,2
Catherine Loudes,3,4
Pierre Gressens,1,2
Annie Faivre-Baumann,3,4
Zsolt Lenkei,5 and
Pascal Dournaud1,2
1Institut National de la Santé et de la Recherche Médicale (INSERM), Unité 676, 75019 Paris, France, 2Université Denis Diderot–Paris 7, 75018 Paris, France, 3INSERM, Unité 549, 75014 Paris, France, 4Université René Descartes–Paris 5, 75006 Paris, France, and 5Ecole Supérieure de Physique et de Chimie Industrielles–Unité Mixte de Recherche Centre National de la Recherche Scientifique 7637, 75005 Paris, France
Correspondence should be addressed to Dr. Pascal Dournaud, Institut National de la Santé et de la Recherche Médicale Unité 676, Hôpital Robert Debré, 48 Boulevard Sérurier, 75019 Paris, France. Email: pascal.dournaud{at}inserm.fr
Despite the large number of G-protein-coupled receptor (GPCR) types expressed in the CNS, little is known about their dynamics in neuronal cells. Dynamic properties of the somatostatin type 2A receptor were therefore examined in resting conditions and after agonist activation in living hippocampal neurons. Using fluorescence recovery after photobleaching experiments, we found that, in absence of ligand, the sst2A receptor is mobile and laterally and rapidly diffuse in neuronal membranes. We then observed by live-cell imaging that, after agonist activation, membrane-associated receptors induce the recruitment of β-arrestin 1–enhanced green fluorescent protein (EGFP) and β-arrestin 2-EGFP to the plasma membrane. In addition, β-arrestin 1-EGFP translocate to the nucleus, suggesting that this protein could serve as a nuclear messenger for the sst2A receptor in neurons. Receptors are then recruited to preexisting clathrin coated pits, form clusters that internalize, fuse, and move to a perinuclear compartment that we identified as the trans-Golgi network (TGN), and recycle. Receptor cargoes are transported through a microtubule-dependent process directly from early endosomes/recycling endosomes to the TGN, bypassing the late endosomal compartment. Together, these results provide a comprehensive description of GPCR trafficking in living neurons and provide compelling evidence that GPCR cargoes can recycle through the TGN after endocytosis, a phenomenon that has not been anticipated from studies of non-neuronal cells.
Key words: β-arrestin; clathrin; endocytosis; FRAP; trans-Golgi network; somatostatin receptor
Received Sept. 25, 2007;
revised Feb. 18, 2008;
accepted Feb. 18, 2008.
Correspondence should be addressed to Dr. Pascal Dournaud, Institut National de la Santé et de la Recherche Médicale Unité 676, Hôpital Robert Debré, 48 Boulevard Sérurier, 75019 Paris, France. Email: pascal.dournaud{at}inserm.fr
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