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Contribution of P2Y(1) receptors to ADP signalling in mouse spinal cord cultures

Amelia E Atterbury-Thomas1, Catherine Leon, Christian Gachet

  • 1Department of Cell Physiology & Pharmacology, Henry Wellcome Building, University of Leicester, Leicester LE1 9HN, UK.

Neuroscience Letters
|March 18, 2008
PubMed

Insights

Adenosine diphosphate (ADP) activates P2Y(1) receptors in spinal cord cultures, increasing intracellular calcium. However, ADP does not stimulate extracellular signal-regulated kinase (ERK) phosphorylation via P2Y(1) receptors in these cells.

Area of Science:

  • Neuroscience
  • Cellular Biology
  • Receptor Pharmacology

Background:

  • Spinal cord neurons and glial cells express purinergic receptors, including P2Y receptors sensitive to adenosine diphosphate (ADP).
  • The specific roles of different P2Y receptor subtypes in spinal cord cellular signaling remain incompletely understood.

Purpose of the Study:

  • To investigate the specific P2Y receptor subtypes involved in ADP-mediated intracellular calcium increases and extracellular signal-regulated protein kinase (ERK) phosphorylation in neonatal mouse spinal cord cultures.
  • To elucidate the signaling pathways activated by ADP in the spinal cord.

Main Methods:

  • Primary mixed neuronal and glial cell cultures were established from neonatal mouse spinal cords.
  • Intracellular calcium levels were measured using fluorescent indicators in response to ADP and UTP.
  • Extracellular signal-regulated protein kinase (ERK) phosphorylation was assessed via Western blotting.
  • Experiments were conducted using wild-type and P2Y(1) receptor-deficient mice, along with specific P2Y(1) receptor antagonists (MRS2179).

Main Results:

  • ADP (10µM) significantly increased intracellular calcium in spinal cord cultures.
  • This ADP-evoked calcium increase was abolished by the P2Y(1) receptor antagonist MRS2179 (10µM) and absent in P2Y(1) receptor-deficient mice.
  • UTP (100µM) evoked calcium increases were unaffected by P2Y(1) receptor deficiency.
  • ADP also robustly increased ERK phosphorylation in both wild-type and P2Y(1) receptor-deficient cultures, indicating a P2Y(1)-independent pathway.

Conclusions:

  • ADP primarily signals through P2Y(1) receptors to elevate intracellular calcium in neonatal spinal cord cultures.
  • The stimulation of ERK phosphorylation by ADP in these cultures occurs independently of P2Y(1) receptors.
  • These findings highlight the differential signaling pathways activated by ADP through specific P2Y receptors in the spinal cord.

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