Endocytosis of Activated Muscarinic m2 Receptor (m2R) in Live Mouse Hippocampal Neurons Occurs via a Clathrin-Dependent Pathway

Lisa Lambert, David Dubayle, Assia Fafouri, Etienne Herzog, Zsolt Csaba, Pascal Dournaud, Salah El Mestikawy, Véronique Bernard
Front. Cell. Neurosci.. 2018-11-30; 12: 450
DOI: 10.3389/fncel.2018.00450

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Lambert L(1), Dubayle D(1)(2), Fafouri A(3), Herzog E(1)(4)(5), Csaba Z(3),
Dournaud P(3), El Mestikawy S(1)(6), Bernard V(1).

Author information:
(1)Sorbonne Université, Université Pierre et Marie Curie UM 119 – CNRS UMR 8246 –
INSERM U1130, Neurosciences Paris Seine – Institut de Biologie Paris Seine (NPS –
IBPS), Paris, France.
(2)Université Paris Descartes – CNRS UMR 8119, Centre de Neurophysique,
Physiologie et Pathologie, Paris, France.
(3)PROTECT, INSERM U1141, Université Paris Diderot, Sorbonne Paris Cité, Paris,
(4)Interdisciplinary Institute for Neuroscience, University Bordeaux, UMR 5297,
Bordeaux, France.
(5)Interdisciplinary Institute for Neuroscience, CNRS, UMR 5297, Bordeaux,
(6)Department of Psychiatry, Douglas Hospital Research Center, McGill University,
Montréal, QC, Canada.

Our aim was to examine the dynamics of the muscarinic m2 receptor (m2R), a
G-protein coupled receptor (GPCR), after agonist activation in living hippocampal
neurons, and especially clathrin dependency endocytosis. We have previously shown
that the m2R undergoes agonist-induced internalization in vivo. However, the
nature of the endocytotic pathway used by m2R after activation is still unknown
in living neurons. Using live cell imaging and quantitative analyses, we have
monitored the effect of stimulation on the fate of the membrane-bound m2R and on
its redistribution in intraneuronal compartments. Shortly (6 min) after
activation, m2R is internalized into clathrin immunopositive structures.
Furthermore, after clathrin-dependent endocytosis, m2R associates with early and
late endosomes and with subcellular organelles involved in degradation. Together,
these results provide, for the first time, a description of m2R trafficking in
living neurons and prove that m2R undergoes clathrin-dependent endocytosis before
being degraded.

DOI: 10.3389/fncel.2018.00450
PMCID: PMC6283979
PMID: 30555302

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