Extracellular transglutaminase-2, nude or associated with astrocytic extracellular vesicles, modulates neuronal calcium homeostasis

Abstract

We have uncovered a novel role for astrocytes-derived extracellular vesicles (EVs) in controlling intraneuronal Ca²⁺ concentration ([Ca²⁺]_i) and identified transglutaminase-2 (TG2) as a surface-cargo of astrocytes-derived EVs. Incubation of hippocampal neurons with primed astrocyte-derived EVs have led to an increase in [Ca²⁺]_i, unlike EVs from TG2-knockout astrocytes. Exposure of neurons or brain slices to extracellular TG2 promoted a [Ca²⁺]_i rise, which was reversible upon TG2 removal and was dependent on Ca²⁺ influx through the plasma membrane. Patch-clamp and calcium imaging recordings revealed TG2-dependent neuronal membrane depolarization and activation of inward currents, due to the Na⁺/Ca²⁺-exchanger (NCX) operating in the reverse mode and indirect activation of L-type VOCCs, as indicated by VOCCs/NCX pharmacological inhibitors. A subunit of Na⁺/K⁺-ATPase was selected by comparative proteomics and identified as being functionally inhibited by extracellular TG2, implicating Na⁺/K⁺-ATPase inhibition in NCX reverse mode-switching leading to Ca²⁺ influx and higher basal [Ca²⁺]_i. These data suggest that reactive astrocytes control intraneuronal [Ca²⁺]_i through release of EVs with TG2 as responsible cargo, which could have a significant impact on synaptic activity in brain inflammation.