A sublethal ischemic episode [termed preconditioning (PC)] protects neurons in the brain against a subsequent severe ischemic injury. This phenomenon is known as brain ischemic tolerance and has received much attention from researchers because of its robust neuroprotective effects. We have previously reported that PC activates astrocytes and subsequently upregulates P2X7 receptors, thereby leading to ischemic tolerance. However, the downstream signals of P2X7 receptors that are responsible for PC-induced ischemic tolerance remain unknown. Here, we show that PC-induced P2X7 receptor-mediated lactate release from astrocytes has an indispensable role in this event. Using a transient focal cerebral ischemia model caused by middle cerebral artery occlusion, extracellular lactate levels during severe ischemia were significantly increased in mice who experienced PC; this increase was dependent on P2X7 receptors. In addition, the intracerebroventricular injection of lactate protected against cerebral ischemic injury. In in vitro experiments, although stimulation of astrocytes with the P2X7 receptor agonist BzATP had no effect on the protein levels of monocarboxylate transporter (MCT) 1 and MCT4 (which are responsible for lactate release from astrocytes), BzATP induced the plasma membrane translocation of these MCTs via their chaperone CD147. Importantly, CD147 was increased in activated astrocytes after PC, and CD147-blocking antibody abolished the PC-induced facilitation of astrocytic lactate release and ischemic tolerance. Taken together, our findings suggest that astrocytes induce ischemic tolerance via P2X7 receptor-mediated lactate release.
Astrocytes , Ischemic Preconditioning , Lactic Acid , Mice, Inbred C57BL , Monocarboxylic Acid Transporters , Receptors, Purinergic P2X7 , Animals , Astrocytes/metabolism , Astrocytes/drug effects , Ischemic Preconditioning/methods , Lactic Acid/metabolism , Lactic Acid/pharmacology , Receptors, Purinergic P2X7/metabolism , Male , Monocarboxylic Acid Transporters/metabolism , Basigin/metabolism , Brain Ischemia/metabolism , Symporters/metabolism , Infarction, Middle Cerebral Artery/metabolism , Disease Models, Animal , Muscle Proteins/metabolism , Adenosine Triphosphate/metabolism , Adenosine Triphosphate/pharmacology , Mice , Cells, Cultured , Brain/metabolism , Mice, Knockout
Although psychotropic drugs act on neurons and glial cells, how glia respond, and whether glial responses are involved in therapeutic effects are poorly understood. Here, we show that fluoxetine (FLX), an anti-depressant, mediates its anti-depressive effect by increasing the gliotransmission of ATP. FLX increased ATP exocytosis via vesicular nucleotide transporter (VNUT). FLX-induced anti-depressive behavior was decreased in astrocyte-selective VNUT-knockout mice or when VNUT was deleted in mice, but it was increased when astrocyte-selective VNUT was overexpressed in mice. This suggests that VNUT-dependent astrocytic ATP exocytosis has a critical role in the therapeutic effect of FLX. Released ATP and its metabolite adenosine act on P2Y11 and adenosine A2b receptors expressed by astrocytes, causing an increase in brain-derived neurotrophic factor in astrocytes. These findings suggest that in addition to neurons, FLX acts on astrocytes and mediates its therapeutic effects by increasing ATP gliotransmission.
Depression/drug therapy , Fluoxetine/administration & dosage , Nucleotide Transport Proteins/genetics , Receptor, Adenosine A2B/genetics , Receptors, Purinergic P2/genetics , Adenosine Triphosphate/metabolism , Animals , Antidepressive Agents/administration & dosage , Astrocytes/drug effects , Astrocytes/metabolism , Astrocytes/pathology , Depression/genetics , Depression/metabolism , Depression/pathology , Exocytosis/drug effects , Gene Expression Regulation/drug effects , Humans , Mice , Mice, Knockout , Neuroglia/drug effects , Neuroglia/metabolism , Neurons/drug effects , Neurons/metabolism
