Purinoceptor-mediated calcium signaling in primary neuron-glia trigeminal cultures

Stefania Ceruti, Marta Fumagalli, Giovanni Villa, Claudia Verderio, Maria P. Abbracchio

Research output: Contribution to journalArticle

Abstract

Receptors for extracellular nucleotides (the P2X-calcium channels and the phospholipase C-coupled P2Y receptors) play key roles in pain signaling, but little is known on their function in trigeminal ganglia, whose hyperactivation leads to the development of migraine pain. Here we characterize calcium signaling via P2X3 and P2Y receptors in primary mouse neuron-glia trigeminal cultures. Comparison with intact ganglion showed that, in dissociated cultures, sensory neurons retain, at least in part, their physical relationships with satellite glia. RT-PCR indicated expression of P2X2/P2X3 (confirmed by immunocytochemistry) and of all cloned P2Y receptors. Single-cell calcium imaging with subtype-selective P2-agonists/antagonists revealed presence of functional neuronal P2X3, as well as of ADP-sensitive P2Y1,12,13 and UTP-activated P2Y2/P2Y4 receptors on both neurons and glia. Calcium responses were much higher in glia, that also responded to UDP, suggesting functional P2Y6 receptors. To study whether trigeminal ganglia P2 receptors are modulated upon treatment with pro-inflammatory agents, cultures were acutely (up to 3 min) or chronically (24 h) exposed to bradykinin. This resulted in potentiation of algogenic P2X3 receptor-mediated calcium responses followed by their down-regulation at 24 h. At this exposure time, P2Y receptors responses in satellite glia were instead upregulated, suggesting a complex modulation of P2 receptors in pain signaling.

Original languageEnglish
Pages (from-to)576-590
Number of pages15
JournalCell Calcium
Volume43
Issue number6
DOIs
Publication statusPublished - Jun 2008

Keywords

  • Bradykinin
  • Calcium signaling
  • P2 purinoceptors
  • Satellite glial cells

ASJC Scopus subject areas

  • Cell Biology
  • Endocrinology

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