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      Resensitization of P2Y receptors by growth factor-mediated activation of the phosphatidylinositol-3 kinase in retinal glial cells.

      Investigative ophthalmology & visual science
      Adenosine Triphosphate, pharmacology, Animals, Calcium, metabolism, Cells, Cultured, Enzyme Activation, Growth Substances, Guinea Pigs, Neuroglia, drug effects, enzymology, Phosphatidylinositol 3-Kinases, Phosphoprotein Phosphatases, Receptors, Purinergic P2, Retina, cytology

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          Abstract

          To determine whether activation of receptor tyrosine kinases enhances the responsiveness of purinergic P2Y receptors in Muller glial cells, known to induce Muller cell proliferation. The P2Y receptors of primary cultured Muller cells of the guinea pig were desensitized by short (30 seconds to 10 minutes)- and long (24 or 48 hours)-term application of adenosine 5'-triphosphate (ATP). Recordings of ATP-evoked intracellular calcium responses showed whether short-term application of different growth factors evoke a resensitization of the receptors. Coapplication of pharmacologic inhibitors showed whether activation of protein kinases is involved in receptor resensitization. Both short- and long-term incubation with ATP induced a significant P2Y receptor desensitization, which was indicated by a strongly reduced intracellular calcium mobilization and which lasted for at least 48 hours. However, the receptors were significantly resensitized after short-term application of platelet-derived, epidermal, or nerve growth factors. The growth factor-mediated resensitization was dependent on an intact cytoskeleton and on the activation of protein phosphatases and of the phosphatidylinositol-3 kinase (PI3K), but was independent of the activation of protein kinase C, src kinases, or extracellular signal-regulated kinases. The results show that activation of receptor tyrosine kinases causes, via activation of PI3K and protein phosphatases, a resensitization of P2Y receptors formerly desensitized by agonist application. The growth factor-mediated resensitization may underlie the previously observed enhanced responsiveness of P2Y receptors in retinal glial cells in experimental retinal detachment and proliferative vitreoretinopathy and may contribute to the induction of reactive gliosis and Muller cell proliferation.

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