Role of the metabotropic P2Y(4) receptor during hypoglycemia: cross talk with the ionotropic NMDAR1 receptor

Exp Cell Res. 2004 Oct 15;300(1):149-58. doi: 10.1016/j.yexcr.2004.07.009.

Abstract

It is well established that both extracellular ATP and glutamate exert a critical role during metabolic impairment, that several P2 receptor subunits are directly involved in this action and that a strong relationship exists between glutamatergic and purinergic signals. Therefore, here we studied the molecular behavior of the purinergic metabotropic P2Y(4) and the glutamatergic ionotropic NMDAR1 receptors during hypoglycemic cell death. We find that these proteins are oppositely modulated during glucose starvation (P2Y(4) is induced, whereas NMDAR1 is inhibited) and that both P2 and NMDA antagonists can restore basal protein expression levels. Moreover, double immunofluorescence experiments with confocal laser microscopy reveal co-localization at the membrane level between the P2Y(4) and NMDAR1 receptors, in both homologous (cerebellar granule neurons) and heterologous (Hek-293) cellular systems. This is furthermore confirmed by co-immunoprecipitation experiments. Finally, when we express the P2Y(4) receptor in the heterologous SH-SY5Y neuronal cell line, hypoglycemia then causes severe cell death and simultaneous downregulation of the NMDAR1 protein. In summary, our work establishes a potential molecular interplay between P2Y(4) and NMDAR1 receptors during glucose deprivation and the causative role of the P2Y(4) during cell death.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / metabolism
  • Animals
  • Animals, Newborn
  • Cell Death / drug effects
  • Cell Death / genetics
  • Cell Line, Tumor
  • Cell Membrane / drug effects
  • Cell Membrane / genetics
  • Cell Membrane / metabolism
  • Cells, Cultured
  • Dizocilpine Maleate / pharmacology
  • Down-Regulation / drug effects
  • Down-Regulation / genetics
  • Epithelial Cells / drug effects
  • Epithelial Cells / metabolism
  • Excitatory Amino Acid Antagonists / pharmacology
  • Glucose / deficiency
  • Glucose / metabolism*
  • Glutamic Acid / metabolism
  • Humans
  • Hypoglycemia / genetics
  • Hypoglycemia / metabolism*
  • Neurons / drug effects
  • Neurons / metabolism
  • Purinergic P2 Receptor Antagonists
  • Rats
  • Rats, Wistar
  • Receptor Cross-Talk / physiology*
  • Receptors, N-Methyl-D-Aspartate / antagonists & inhibitors
  • Receptors, N-Methyl-D-Aspartate / genetics
  • Receptors, N-Methyl-D-Aspartate / metabolism*
  • Receptors, Purinergic P2 / genetics
  • Receptors, Purinergic P2 / metabolism*
  • Signal Transduction / physiology
  • Triazines / pharmacology

Substances

  • Excitatory Amino Acid Antagonists
  • NMDA receptor A1
  • Purinergic P2 Receptor Antagonists
  • Receptors, N-Methyl-D-Aspartate
  • Receptors, Purinergic P2
  • Triazines
  • purinoceptor P2Y4
  • Glutamic Acid
  • Cibacron Blue F 3GA
  • Dizocilpine Maleate
  • Adenosine Triphosphate
  • Glucose