Extracellular ATP-prinoceptor signaling and AMP-activated protein kinase regulate astrocytic glucose transporter 3 in an in vitro ischemia

Astrocytes become hypertrophic reactive in response to the ischemic stress, and they contribute to either protect or exacerbate neuronal damage, depending on the depth or duration of the stress. Astrocytes have more resistance to the ischemic stress than neurons, which is apparently due to active anerobic metabolic pathway in the emergency situation. We have been focused on the functional role of astrocytic glucose transporters in the ischemic condition. Under the physiological conditions, cultured astrocytes primarily express glucose transporter1 (GLUT1), and GLUT3 is only detected at extremely low levels. But astrocytes enhance GLUT3 expression through the signaling of nuclear factor-κ-light-chain-enhancer of activated B cells (NF-κB) under mild ischemic condition. It is reasonable since GLUT3 transports extracellular glucose about seven times faster than GLUT1, so astrocytes enhance the storage of intracellular glucose during the ischemia. However, other signaling cascades that regulate GLUT3 production remain unknown. Here we demonstrate that extracellular adenosine 5'-triphosphate (ATP)-P2Y receptor signaling also regulates GLUT3 expression. Under mild ischemic condition, astrocytes positively released existing intracellular or newly synthesized ATP by AMP-activated protein kinase (AMPK) signaling. The released extracellular ATP from pore channels activated ATP-sensitive P2Y receptor signaling, resulting in an increase in c-Fos and c-Jun proteins. Newly synthesized GLUT3 was regulated by those signaling since the inhibition of P2Y receptors or c-Fos/c-Jun signaling significantly reduced GLUT3 expression. Furthermore, the inhibition of P2Y receptors during the ischemic condition sustained intracellular ATP concentration, leading to a decrease in AMPK proteins. These results suggest AMPK-regulated ATP production triggers the release of ATP to activate P2Y receptor signaling, which is another candidate that regulates GLUT3 expression under the ischemic condition.