Traumatic injury activates protein kinase B/Akt in cultured astrocytes: role of extracellular ATP and P2 purinergic receptors.
Protein kinase B/Akt is a key signaling molecule that regulates cell survival, growth, and metabolism, and inhibits apoptosis. Traumatic brain injury (TBI) activates Akt, and Akt has been implicated in neuronal survival after TBI, but little is known about injury-induced Akt activation in astrocytes, cells that exhibit hypertrophic and hyperplastic responses to CNS injury. Here we have investigated the effect of mechanical strain on Akt activation in primary cultures of rat cortical astrocytes growing on deformable Silastic membranes. When astrocytes were subjected to mechanical strain (50 msec; 5-7.5 mm displacement), we observed an increase in phosphorylation of serine 473, a key indicator of Akt activation. Akt phosphorylation was increased at 3 min postinjury, was maximal from 5 to 10 min, and declined gradually thereafter. Akt activation was also dependent on the severity of the injury. Stretch-induced Akt phosphorylation was attenuated by blocking calcium influx and phosphoinositide 3-kinase (PI3K), an upstream activator of Akt. In addition, we found that ATP is rapidly released after mechanical strain and that the P2 purinergic receptor antagonist iso-pyridoxal-5'-phosphate-6-azophenyl-2',5'disulfonate (PPADS) attenuated trauma-induced Akt activation. We conclude that mechanical strain causes activation of Akt in astrocytes via stimulation of P2 receptors. This suggests that P2 receptor/Akt signaling promotes astrocyte survival and growth, and this process may play a role in the generation of reactive gliosis after TBI.
Neary
Joseph T
JT
Research Service, VA Medical Center, Departments of Pathology, Biochemistry and Molecular Biology and Neuroscience Program, University of Miami School of Medicine, Miami, Florida.
Kang
Yuan
Y
Tran
Minh
M
Feld
Jonathan
J
D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000201:enzymology / Q000378:metabolism*; D001930:Brain Injuries / Q000378:metabolism* / Q000503:physiopathology; D015220:Calcium Channels / Q000187:drug effects / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000293:injuries / Q000378:metabolism / Q000503:physiopathology; D045604:Extracellular Fluid / Q000378:metabolism; D005911:Gliosis / Q000201:enzymology / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D000081082:Phosphoinositide-3 Kinase Inhibitors; D010766:Phosphorylation; D010975:Platelet Aggregation Inhibitors / Q000494:pharmacology; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D051057:Proto-Oncogene Proteins c-akt; D058919:Purinergic P2 Receptor Antagonists; D011732:Pyridoxal Phosphate / Q000031:analogs & derivatives* / Q000494:pharmacology; D051381:Rats; D018048:Receptors, Purinergic P2 / Q000378:metabolism*; D012694:Serine / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D013314:Stress, Mechanical
D000255:Adenosine Triphosphate; D015220:Calcium Channels; D019869:Phosphatidylinositol 3-Kinases; D000081082:Phosphoinositide-3 Kinase Inhibitors; D010975:Platelet Aggregation Inhibitors; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins; D051057:Proto-Oncogene Proteins c-akt; D058919:Purinergic P2 Receptor Antagonists; D011732:Pyridoxal Phosphate; D018048:Receptors, Purinergic P2; D012694:Serine; C077792:pyridoxal phosphate-6-azophenyl-2',4'-disulfonic acid; C494920:Akt1 protein, rat