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The Journal of Neuroscience, November 15, 2002, 22(22):9776-9784

Rab11a and Myosin Vb Regulate Recycling of the M4 Muscarinic Acetylcholine Receptor

Laura A. Volpicelli1, James J. Lah1, Guofu Fang1, James R. Goldenring2, and Allan I. Levey1

1 Center for Neurodegenerative Disease and Department of Neurology, Emory University School of Medicine, Atlanta, Georgia 30322, and 2 Department of Surgery, Vanderbilt University School of Medicine, Nashville, Tennessee 37232

Agonist-induced internalization followed by subsequent return to the cell surface regulates G-protein-coupled receptor (GPCR) activity. Because the cellular responsiveness to ligand depends on the balance between receptor degradation and recycling, it is crucial to identify the molecules involved in GPCR recovery to the cell surface. In this study, we identify mechanisms involved in the recycling of the M4 subtype of muscarinic acetylcholine receptor. M4 is highly expressed in the CNS, plays a role in locomotor activity, and is a novel therapeutic target for neurologic and psychiatric disorders. Previous studies show that, after cholinergic stimulation, M4 internalizes from the cell surface to endosomes in cell culture and the rat brain. Here, we show that, after activation, M4 traffics to transferrin receptor- and Rab11a-positive perinuclear endosomes. Expression of the constitutively GDP-bound, inactive mutant Rab11aS25N inhibits M4 trafficking to recycling endosomes. Expression of the C-terminal tail of myosin Vb, a Rab11a effector, enhances M4 accumulation in perinuclear endosomes. Both Rab11aS25N and the myosin Vb tail impair M4 recycling. The results demonstrate that GPCR recycling is mediated through a discrete pathway using both Rab11a and myosin Vb.

Key words: PC12 cells; muscarinic acetylcholine receptors; G-protein-coupled receptor; M4; recycling; endosomes; Rab11a; myosin Vb; unconventional myosins; transferrin receptor; endocytosis; internalization


Copyright © 2002 Society for Neuroscience  0270-6474/02/22229776-09$05.00/0


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