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Cited by 126Adenosine and Purinergic SignalingIon Channels and ReceptorsCalcium signaling and nucleotide metabolism

Properties of the pore‐forming P2X7 purinoceptor in mouse NTW8 microglial cells

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Chessell, I P, Michel, A D, Humphrey, P P A·1997·British Journal of Pharmacology
DOI 10.1038/sj.bjp.0701278PMC PMC1564837
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Abstract

We have used whole‐cell patch clamping methods to study and characterize the cytolytic P2X 7 (P2 Z) receptor in the NTW8 mouse microglial cell line. At room temperature, in an extracellular solution containing 2 m M Ca 2+ and 1 m M Mg 2+, 2′‐ and 3′‐O‐(4‐benzoylbenzoyl)‐adenosine‐5′‐triphosphate (Bz‐ATP; 300 μ M), or ATP (3 m M), evoked peak whole cell inward currents, at a holding potential of −90 mV, of 549±191 and 644±198 pA, respectively. Current‐voltage relationships generated with 3 m M ATP reversed at 4.6 mV and did not display strong rectification. In an extracellular solution containing zero Mg 2+ and 500 μ M Ca 2+ (low divalent solution), brief (0.5 s) application of these agonists elicited larger maximal currents (909±138 and 1818±218 pA, Bz‐ATP and ATP, respectively). Longer application of ATP (1 m M for 30 s) produced larger, slowly developing, currents which reached a plateau after approximately 15–20 s and were reversible on washing. Under these conditions, in the presence of ATP, ethidium bromide uptake could be demonstrated. Further applictions of 1 m M ATP produced rapid currents of the same magnitude as those observed during the 30 s application. Subsequent determination of concentration‐effect curves to Bz‐ATP, ATP and 2‐methylthio‐ATP yielded EC 50 values of 58.3, 298 and 505 μ M, respectively. These affects of ATP were antagonized by pyridoxal‐phosphate‐6‐azophenyl‐ 2′, 4′‐disulphonic acid (PPADS; 30 μ M) but not suramin (100 μ M). In low divalent solution, repeated application of 1 m M ATP for 1 s produced successively larger currents which reached a plateau, after 8 applications, of 466% of the first application current. PPADS (30 μ M) prevented this augmentation, while 5‐(N,N‐hexamethylene)‐amiloride (HMA) (100 μ M) accelerated it such that maximal augmentation was observed after only one application of ATP in the presence of HMA. At a bath temperature of 32°C, current augmentation also occurred in normal divalent cation containing solution. These data demonstrate that mouse microglial NTW8 cells possess a purinoceptor with pharmacological characteristics resembling the P2X 7 receptor. We suggest that the current augmentation phenomenon observed reflects formation of the large cytolytic pore characteristic of this receptor. We have demonstrated that pore formation can occur under normal physiological conditions and can be modulated pharmacologically, both positively and negatively.

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  • The P2X7 receptor is a trimeric ionotropic receptor with low affinity for ATP (EC50 around 0.3-1.8 mM) and, under prolonged stimulation, can form pores allowing the passage of large hydrophilic molecules.

    “The P2X7 receptor is a trimeric ionotropic receptor that belongs to the P2X family of purinergic receptors, presenting low affinity for ATP (EC50 around 0.3–1.8 mM) and, under prolonged stimulation, can form pores allowing the passage of large hydrophilic molecules [62–64].”

    From Hyperactivation of P2X7 receptors as a culprit of COVID-19 neuropathology (2021) · cites this paper as [62]

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Authors
Chessell, I. P., Michel, A. D., & Humphrey, P. P. A.
Affiliations
University of Cambridge (GB)
Corresponding author
Chessell, I P
Venue
British Journal of Pharmacology
Publisher
Wiley
Published
1997-07
Volume
121
Issue
7
Pages
1429-1437
ISSN
  • 0007-1188 (ISSN-L)
  • 1476-5381
DOI
10.1038/sj.bjp.0701278
PMC
PMC1564837
OpenAlex
W1989632372
Cited by
126
References
30
Citations by year
2024: 2 · 2023: 3 · 2022: 1 · 2021: 3 · 2020: 4 · 2018: 3 · 2017: 2 · 2016: 3 · 2015: 1 · 2013: 7 · 2012: 1
FWCI
3.82
Type
journal article
Language
en
Open access
Licence
onlinelibrary.wiley.com/termsAndConditions#vor (vor, 4242 d)
Locations
  • British Journal of Pharmacology (publishedVersion)
  • Europe PMC (PubMed Central) (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Physiology › Adenosine and Purinergic Signaling
  • Life Sciences › Neuroscience › Sensory Systems › Ion Channels and Receptors
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Physiology › Calcium signaling and nucleotide metabolism
Keywords
P2X7 receptor, microglia, ATP, 2-methylthio-ATP, PPADS, amiloride, divalent cations, magnesium, receptor desensitization, receptor resensitization, cell lysis, purinergic signaling
MeSH terms
Adenosine Triphosphate, Amiloride, Animals, Cell Line, Ethidium, Microglia, Receptors, Purinergic P2, Mice

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