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Expression of P2X receptors in LAD-2 cells and human lung mast cells [Abstract]

Timmermans, J. P., De, Proost, Pintelon, I., Wilkinson, William J., Goethals, S., Brouns, I., Van Nassauw, L., Riccardi, Daniela, Kemp, Paul J. and Adriaensen, D. 2008. Expression of P2X receptors in LAD-2 cells and human lung mast cells [Abstract]. Purinergic Signalling 4 (S1) , S159. 10.1007/s11302-008-9116-0

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Abstract

Mast cells play an important role in the pathophysiology of many allergic diseases including asthma. This is via the release of mediators into surrounding tissues following a rise in intracellular calcium. The molecular identity of the calcium channels supporting this influx have not yet been fully elucidated. P2X receptors are non-selective cation channels which may contribute to the influx of extracellular calcium in mast cells. The purpose of this study was to determine if P2X purinoceptors are functionally present in the LAD-2 mast cell line and primary human lung mast cells (HLMC’s). RT-PCR revealed P2X1, 4 and 7 transcripts in both cell types; mRNA for P2X6 was also detected in LAD-2 cells. Under wholecell patch clamp recording conditions, ATP (1 s application) consistently evoked a P2X1-like current in both cell types. This current could also be elicited by alphabetamethyleneATP (10 microM, 93.55%cells, n=31) and was antagonised in LAD-2 cells by NF 449 (1microM) and Pyridoxal phosphate-6-azo(benzene-2,4-disulfonic acid) tetrasodium salt (10 microM). A P2X7 -like non-desensitising current in response to high concentrations of ATP (1–5 mM) was also seen in both cell types (96% LAD-2 cells, n=24; 54% HLMC’s, n=24) and could be evoked in LAD-2 cells by the agonist 2′(3′)-0-(4-benzoylbenzoyl)ATP (300microM). A P2X4 like current was seen in response to 100microM ATP (80% LAD-2 cells, n=10; 21% HLMC’s, n=29), the amplitude and duration of which was potentiated by Ivermectin. In summary, the electrophysiological recordings support the molecular studies and confirm the presence of functional P2X1, 4 and 7 receptors in LAD-2 and HLMC’s.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Biosciences
Additional Information: Purines 2008 Meeting 29 June – 2 July 2008, Copenhagen, Denmark
Publisher: Springer
ISSN: 1573-9538
Last Modified: 04 Jun 2017 02:07
URI: http://orca-mwe.cf.ac.uk/id/eprint/9285

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