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J Neurosci ; 28(17): 4336-49, 2008 Apr 23.
Artigo em Inglês | MEDLINE | ID: mdl-18434512

RESUMO

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 sst(2A) 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 beta-arrestin 1-enhanced green fluorescent protein (EGFP) and beta-arrestin 2-EGFP to the plasma membrane. In addition, beta-arrestin 1-EGFP translocate to the nucleus, suggesting that this protein could serve as a nuclear messenger for the sst(2A) 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.


Assuntos
Hipocampo/metabolismo , Neurônios/metabolismo , Receptores de Somatostatina/metabolismo , Animais , Células Cultivadas , Difusão , Hipocampo/citologia , Hipocampo/fisiologia , Humanos , Camundongos , Neurônios/citologia , Neurônios/fisiologia , Transporte Proteico/fisiologia , Ratos , Ratos Sprague-Dawley , Receptores de Somatostatina/fisiologia
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