The cytokine IL-1beta transiently enhances P2X7 receptor expression and function in human astrocytes

Glia. 2005 Jan 15;49(2):245-58. doi: 10.1002/glia.20110.

Abstract

Extracellular nucleotide di- and triphosphates such as ATP and ADP mediate their effects through purinergic P2 receptors belonging to either the metabotropic P2Y or the ionotropic P2X receptor family. The P2X7R is a unique member of the P2X family, which forms a pore in response to ligand stimulation, regulating cell permeability, cytokine release, and/or apoptosis. This receptor is also unique in that its affinity for the ligand benzoyl-benzoyl ATP (BzATP) is at least 10-fold greater than that of ATP. Primary human fetal astrocytes in culture express low-levels of P2X7R mRNA and protein, and BzATP induces only a slight influx in intracellular calcium [Ca2+]i, with little demonstrable effect on gene expression or pore formation in these cells. We now show that, following treatment with the proinflammatory cytokine IL-1beta, BzATP induces a robust rise in [Ca2+]i with agonist and antagonist profiles indicative of the P2X7R. IL-1beta also induced the formation of membrane pores as evidenced by the uptake of YO-PRO-1 (375 Da). Quantitative real-time PCR demonstrated transient upregulation of P2X7R mRNA in IL-1beta-treated cells, while FACS analysis indicated a similar upregulation of P2X7R protein at the cell membrane. In multiple sclerosis lesions, immunoreactivity for the P2X7R was demonstrated on reactive astrocytes in autopsy brain tissues. In turn, P2X7R stimulation increased the production of IL-1-induced nitric oxide synthase activity by astrocytes in culture. These studies suggest that signaling via the P2X7R may modulate the astrocytic response to inflammation in the human central nervous system.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenosine Triphosphate / analogs & derivatives*
  • Adenosine Triphosphate / pharmacology
  • Astrocytes / drug effects
  • Astrocytes / metabolism*
  • Benzoxazoles
  • Calcium Signaling / drug effects
  • Calcium Signaling / physiology
  • Cell Membrane / drug effects
  • Cell Membrane / genetics
  • Cell Membrane / metabolism
  • Cells, Cultured
  • Encephalitis / genetics
  • Encephalitis / metabolism*
  • Fluorescent Dyes / pharmacokinetics
  • Humans
  • Interleukin-1 / metabolism*
  • Interleukin-1 / pharmacology
  • Multiple Sclerosis / genetics
  • Multiple Sclerosis / metabolism
  • Nitric Oxide / biosynthesis
  • Protein Transport / drug effects
  • Protein Transport / genetics
  • Quinolinium Compounds
  • RNA, Messenger / drug effects
  • RNA, Messenger / metabolism
  • Receptors, Purinergic P2 / drug effects
  • Receptors, Purinergic P2 / genetics
  • Receptors, Purinergic P2 / metabolism*
  • Receptors, Purinergic P2X7
  • Signal Transduction / drug effects
  • Signal Transduction / genetics
  • Up-Regulation / drug effects
  • Up-Regulation / genetics

Substances

  • Benzoxazoles
  • Fluorescent Dyes
  • Interleukin-1
  • P2RX7 protein, human
  • Quinolinium Compounds
  • RNA, Messenger
  • Receptors, Purinergic P2
  • Receptors, Purinergic P2X7
  • YO-PRO 1
  • Nitric Oxide
  • 3'-O-(4-benzoyl)benzoyladenosine 5'-triphosphate
  • Adenosine Triphosphate