Constitutive expression of CCR2 chemokine receptor and inhibition by MCP-1/CCL2 of GABA-induced currents in spinal cord neurones

J Neurochem. 2005 Nov;95(4):1023-34. doi: 10.1111/j.1471-4159.2005.03431.x. Epub 2005 Sep 7.

Abstract

In the CNS, immune-like competent cells (microglia and astrocytes) were first described as potential sites of chemokine synthesis, but more recent evidence has indicated that neurones might also express chemokines and their receptors. The aim of the present work was to investigate further, both in vivo and in vitro, CC Chemokine Family Receptor 2 (CCR2) expression and functionality in rat spinal cord neurones. First, we demonstrated by RT-PCR and western blot analysis that CCR2 mRNA and protein were present in spinal extracts. Furthermore, we showed by immunolabelling that CCR2 was exclusively expressed by neurones in spinal sections of healthy rat. Finally, to test the functionality of CCR2, we used primary cultures of rat spinal neurones. In this model, similar to what was observed in vivo, CCR2 mRNA and protein were expressed by neurones. Cultured neurones stimulated with Monocyte Chemoattractant Protein-1 (MCP-1)/CCL2, the best characterized CCR2 agonist, showed activation of the Akt pathway. Finally, patch-clamp recording of cultured spinal neurones was used to investigate whether MCP-1/CCL2 could modulate their electrophysiological properties. MCP-1 alone did not affect the electrical properties of spinal neurones, but potently and efficiently inhibited GABA(A)-mediated GABAergic responses in these neurones. These data constitute the first demonstration of a modulatory role of MCP-1 on GABAergic neurotransmission and contribute to our understanding of the roles of CCR2 and MCP-1/CCL2 in spinal cord physiology, in particular with respect to nociceptive transmission, as well as the implication of this chemokine in neuronal adaptation or dysfunction during neuropathy.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Autoradiography / methods
  • Bicuculline / pharmacology
  • Blotting, Northern / methods
  • Blotting, Western / methods
  • Cells, Cultured
  • Chemokine CCL2 / pharmacology*
  • Dose-Response Relationship, Drug
  • Drug Interactions
  • ELAV Proteins / metabolism
  • Embryo, Mammalian
  • Female
  • GABA Antagonists / pharmacology
  • Gene Expression Regulation / drug effects*
  • Gene Expression Regulation / physiology
  • Glial Fibrillary Acidic Protein / metabolism
  • Immunohistochemistry / methods
  • Male
  • Membrane Potentials / drug effects
  • Membrane Potentials / physiology
  • Neurons / drug effects*
  • Neurons / physiology
  • Oncogene Protein v-akt / metabolism
  • Patch-Clamp Techniques / methods
  • Phosphorylation
  • Pregnancy
  • RNA, Messenger / metabolism
  • Rats
  • Rats, Wistar
  • Receptors, CCR2
  • Receptors, Chemokine / genetics
  • Receptors, Chemokine / metabolism*
  • Reverse Transcriptase Polymerase Chain Reaction / methods
  • Spinal Cord / cytology*
  • gamma-Aminobutyric Acid / pharmacology*

Substances

  • Ccl2 protein, mouse
  • Ccr2 protein, mouse
  • Ccr2 protein, rat
  • Chemokine CCL2
  • ELAV Proteins
  • GABA Antagonists
  • Glial Fibrillary Acidic Protein
  • RNA, Messenger
  • Receptors, CCR2
  • Receptors, Chemokine
  • gamma-Aminobutyric Acid
  • Oncogene Protein v-akt
  • Bicuculline