Tissue-type plasminogen activator controls neuronal death by raising surface dynamics of extrasynaptic NMDA receptors.
N-methyl-d-aspartate receptors (NMDARs) are ion channels whose synaptic versus extrasynaptic localization critically influences their functions. This distribution of NMDARs is highly dependent on their lateral diffusion at the cell membrane. Each obligatory subunit of NMDARs (GluN1 and GluN2) contains two extracellular clamshell-like domains with an agonist-binding domain and a distal N-terminal domain (NTD). To date, the roles and dynamics of the NTD of the GluN1 subunit in NMDAR allosteric signaling remain poorly understood. Using single nanoparticle tracking in mouse neurons, we demonstrate that the extracellular neuronal protease tissue-type plasminogen activator (tPA), well known to have a role in the synaptic plasticity and neuronal survival, leads to a selective increase of the surface dynamics and subsequent diffusion of extrasynaptic NMDARs. This process explains the previously reported ability of tPA to promote NMDAR-mediated calcium influx. In parallel, we developed a monoclonal antibody capable of specifically blocking the interaction of tPA with the NTD of the GluN1 subunit of NMDAR. Using this original approach, we demonstrate that the tPA binds the NTD of the GluN1 subunit at a lysine in position 178. Accordingly, when applied to mouse neurons, our selected antibody (named Glunomab) leads to a selective reduction of the tPA-mediated surface dynamics of extrasynaptic NMDARs, subsequent signaling and neurotoxicity, both in vitro and in vivo. Altogether, we demonstrate that the tPA is a ligand of the NTD of the obligatory GluN1 subunit of NMDAR acting as a modulator of their dynamic distribution at the neuronal surface and subsequent signaling.
Inserm, Inserm UMR-S U919, Serine Proteases and Pathophysiology of the Neurovascular Unit, Université Caen-Normandie, GIP Cyceron, Caen, France.
Flavie
F
Lesept
Inserm, Inserm UMR-S U919, Serine Proteases and Pathophysiology of the Neurovascular Unit, Université Caen-Normandie, GIP Cyceron, Caen, France.
Arnaud
A
Chevilley
Interdisciplinary Institute for Neuroscience, Université de Bordeaux UMR 5297, Bordeaux, France.|CNRS IINS UMR 5297, Bordeaux, France.
Julie
J
Jezequel
Interdisciplinary Institute for Neuroscience, Université de Bordeaux UMR 5297, Bordeaux, France.|CNRS IINS UMR 5297, Bordeaux, France.
Laurent
L
Ladépêche
Inserm, Inserm UMR-S U919, Serine Proteases and Pathophysiology of the Neurovascular Unit, Université Caen-Normandie, GIP Cyceron, Caen, France.
Richard
R
Macrez
Inserm, Inserm UMR-S U919, Serine Proteases and Pathophysiology of the Neurovascular Unit, Université Caen-Normandie, GIP Cyceron, Caen, France.
Margaux
M
Aimable
Inserm, Inserm UMR-S U919, Serine Proteases and Pathophysiology of the Neurovascular Unit, Université Caen-Normandie, GIP Cyceron, Caen, France.
Sophie
S
Lenoir
Inserm, Inserm UMR-S U919, Serine Proteases and Pathophysiology of the Neurovascular Unit, Université Caen-Normandie, GIP Cyceron, Caen, France.
Thomas
T
Bertrand
SysDiag CNRS/Bio-Rad, UMR3145, Montpellier, France.
Laëtitia
L
Rubrecht
SysDiag CNRS/Bio-Rad, UMR3145, Montpellier, France.
Pascale
P
Galea
Inserm, Inserm UMR-S U919, Serine Proteases and Pathophysiology of the Neurovascular Unit, Université Caen-Normandie, GIP Cyceron, Caen, France.
Laurent
L
Lebouvier
PAION Deutschland GmbH, Aachen, Germany.
Karl-Uwe
KU
Petersen
Inserm, Inserm UMR-S U919, Serine Proteases and Pathophysiology of the Neurovascular Unit, Université Caen-Normandie, GIP Cyceron, Caen, France.
Yannick
Y
Hommet
Inserm, Inserm UMR-S U919, Serine Proteases and Pathophysiology of the Neurovascular Unit, Université Caen-Normandie, GIP Cyceron, Caen, France.
Eric
E
Maubert
Inserm, Inserm UMR-S U919, Serine Proteases and Pathophysiology of the Neurovascular Unit, Université Caen-Normandie, GIP Cyceron, Caen, France.
Carine
C
Ali
Interdisciplinary Institute for Neuroscience, Université de Bordeaux UMR 5297, Bordeaux, France.|CNRS IINS UMR 5297, Bordeaux, France.
Laurent
L
Groc
Inserm, Inserm UMR-S U919, Serine Proteases and Pathophysiology of the Neurovascular Unit, Université Caen-Normandie, GIP Cyceron, Caen, France.
Denis
D
Vivien
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism*; D004058:Diffusion; D005341:Fibrinolysin / Q000494:pharmacology; D057809:HEK293 Cells; D006801:Humans; D008239:Lysine / Q000378:metabolism; D008297:Male; D008807:Mice, Inbred BALB C; D009474:Neurons / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D009498:Neurotoxins / Q000633:toxicity; D000072417:Protein Domains; D017207:Rats, Sprague-Dawley; D016194:Receptors, N-Methyl-D-Aspartate / Q000737:chemistry / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D013569:Synapses / Q000187:drug effects / Q000378:metabolism*; D010959:Tissue Plasminogen Activator / Q000494:pharmacology*
D000911:Antibodies, Monoclonal; D009498:Neurotoxins; D016194:Receptors, N-Methyl-D-Aspartate; D010959:Tissue Plasminogen Activator; D005341:Fibrinolysin; D008239:Lysine; D002118:Calcium