TY - JOUR
T1 - P2X7-dependent, but differentially regulated release of IL-6, CCL2, and TNF-alpha in cultured mouse microglia
AU - Shieh, Chu-Hsin
AU - Heinrich, Annette
AU - Serchov, Tsvetan
AU - van Calker, Dietrich
AU - Biber, Knut
PY - 2014/4
Y1 - 2014/4
N2 - ATP is an important regulator of microglia and its effects on microglial cytokine release are currently discussed as important contributors in a variety of brain diseases. We here analyzed the effects of ATP on the production of six inflammatory mediators (IL-6, IL-10, CCL2, IFN-gamma, TNF-alpha, and IL-12p70) in cultured mouse primary microglia. Stimulation of P2X7 receptor by ATP (1 mM) or BzATP (500 mu M) evoked the mRNA expression and release of proinflammatory cytokines IL-6, TNF-alpha, and the chemokine CCL2 in WT cells but not in P2X7(-/-) cells. The effects of ATP and BzATP were inhibited by the nonselective P2 receptor antagonists PPADs and suramin. Various selective P2X7 receptor antagonists blocked the P2X7-dependent release of IL-6 and CCL2, but, surprisingly, had no effect on BzATP-induced release of TNF-alpha in microglia. Calcium measurements confirmed that P2X7 is the main purine receptor activated by BzATP in microglia and showed that all P2X7 antagonists were functional. It is also presented that pannexin-1 hemichannel function and potential P2X4/P2X7 heterodimers are not involved in P2X7-dependent release of IL-6, CCL2, and TNF-alpha in microglia. How P2X7-specific antagonists only affect P2X7-dependent IL-6 and CCL2 release, but not TNF-alpha release is at the moment unclear, but indicates that the P2X7-dependent release of cytokines in microglia is differentially regulated. GLIA 2014;62:592-607
AB - ATP is an important regulator of microglia and its effects on microglial cytokine release are currently discussed as important contributors in a variety of brain diseases. We here analyzed the effects of ATP on the production of six inflammatory mediators (IL-6, IL-10, CCL2, IFN-gamma, TNF-alpha, and IL-12p70) in cultured mouse primary microglia. Stimulation of P2X7 receptor by ATP (1 mM) or BzATP (500 mu M) evoked the mRNA expression and release of proinflammatory cytokines IL-6, TNF-alpha, and the chemokine CCL2 in WT cells but not in P2X7(-/-) cells. The effects of ATP and BzATP were inhibited by the nonselective P2 receptor antagonists PPADs and suramin. Various selective P2X7 receptor antagonists blocked the P2X7-dependent release of IL-6 and CCL2, but, surprisingly, had no effect on BzATP-induced release of TNF-alpha in microglia. Calcium measurements confirmed that P2X7 is the main purine receptor activated by BzATP in microglia and showed that all P2X7 antagonists were functional. It is also presented that pannexin-1 hemichannel function and potential P2X4/P2X7 heterodimers are not involved in P2X7-dependent release of IL-6, CCL2, and TNF-alpha in microglia. How P2X7-specific antagonists only affect P2X7-dependent IL-6 and CCL2 release, but not TNF-alpha release is at the moment unclear, but indicates that the P2X7-dependent release of cytokines in microglia is differentially regulated. GLIA 2014;62:592-607
KW - neuroinflammation
KW - antagonists
KW - purine receptors
KW - neuroprotection
KW - TUMOR-NECROSIS-FACTOR
KW - EXPERIMENTAL AUTOIMMUNE ENCEPHALOMYELITIS
KW - FOCAL CEREBRAL-ISCHEMIA
KW - P2X(7) RECEPTOR
KW - P2X7 RECEPTOR
KW - EXTRACELLULAR ATP
KW - PURINERGIC RECEPTORS
KW - ACTIVATED MICROGLIA
KW - CYTOKINE RELEASE
KW - INTERLEUKIN-1-BETA RELEASE
U2 - 10.1002/glia.22628
DO - 10.1002/glia.22628
M3 - Article
SN - 0894-1491
VL - 62
SP - 592
EP - 607
JO - Glia
JF - Glia
IS - 4
ER -