title: Recycling endosomes undergo rapid closure of a fusion pore on exocytosis in neuronal dendrites.
abstract: Exocytosis of recycling endosomes (REs) represents the last step of receptor and membrane recycling, a fundamental process involved in many aspects of cell physiology. In neurons, it is involved in the control of cell polarity and synaptic plasticity and is locally and tightly regulated. However, its molecular mechanisms are still poorly understood. We have imaged single exocytosis events of REs in rat hippocampal neurons in culture transfected with three types of receptors tagged with the pH-sensitive GFP mutant superecliptic phluorin. We found that exocytosis events are grouped into two categories: (1) short burst events in which receptors diffuse into the plasma membrane in a few seconds; and (2) long display events in which receptors remain visible and clustered after exocytosis for many seconds. Display events are much rarer in non-neuronal cells, such as fibroblasts and astrocytes. Using two-color imaging and fast extracellular solution changes, we show that display events correspond to the rapid opening and closing of a fusion pore (or "kiss-and-run") with a median opening time of 2.6 s, which restricts the diffusion of multiple receptor types and bound cargo. Moreover, the RE marker Rab11 remains enriched after display exocytosis events and controls the mode of RE exocytosis. Finally, a given RE can undergo multiple rounds of display exocytosis. The last step of recycling can thus be controlled in neurons for the selective delivery of receptors at the cell surface.
authors.affiliation: University of Bordeaux and National Center of Scientific Research, Interdisciplinary Institute for Neuroscience, Coeducational Research Unit 5297, F-33000 Bordeaux, France.
authors.forename: Damien
authors.initials: D
authors.lastname: Jullié
authors.affiliation: University of Bordeaux and National Center of Scientific Research, Interdisciplinary Institute for Neuroscience, Coeducational Research Unit 5297, F-33000 Bordeaux, France.
authors.forename: Daniel
authors.initials: D
authors.lastname: Choquet
authors.affiliation: University of Bordeaux and National Center of Scientific Research, Interdisciplinary Institute for Neuroscience, Coeducational Research Unit 5297, F-33000 Bordeaux, France david.perrais@u-bordeaux.fr.
authors.forename: David
authors.identifier: http://orcid.org/0000-0002-5878-5408
authors.initials: D
authors.lastname: Perrais
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D003712:Dendrites / Q000378:metabolism*; D034281:Dynamins / Q000378:metabolism; D011992:Endosomes / Q000378:metabolism*; D005089:Exocytosis / Q000502:physiology*; D009474:Neurons / Q000378:metabolism*; D051381:Rats
chemical_list: D000199:Actins; D034281:Dynamins
keywords: AMPA receptor; Rab11; astrocyte; beta-adrenergic receptor; kiss-and-run; transferrin receptor