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The Journal of Neuroscience, May 21, 2008, 28(21):5473-5480; doi:10.1523/JNEUROSCI.1149-08.2008

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Cellular/Molecular
P2X₁ and P2X₅ Subunits Form the Functional P2X Receptor in Mouse Cortical Astrocytes

Ulyana Lalo,¹ Yuri Pankratov,^1,2 Sven P. Wichert,³ Moritz J. Rossner,³ R. Alan North,¹ Frank Kirchhoff,³ and Alexei Verkhratsky^1,4

¹Faculty of Life Sciences, The University of Manchester, Manchester M13 9PT, United Kingdom, ²Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, United Kingdom, ³Department of Neurogenetics, Max Planck Institute of Experimental Medicine, D-37075 Göttingen, Germany, and ⁴Institute of Experimental Medicine, Academy of Sciences of the Czech Republic, 142 20 Prague 4, Czech Republic

Correspondence should be addressed to Prof. Alexei Verkhratsky, Faculty of Life Sciences, The University of Manchester, 1.124 Stopford Building, Oxford Road, Manchester M13 9PT, UK. Email: alex.verkhratsky{at}manchester.ac.uk

ATP plays an important role in signal transduction between neuronal and glial circuits and within glial networks. Here we describe currents activated by ATP in astrocytes acutely isolated from cortical brain slices by non-enzymatic mechanical dissociation. Brain slices were prepared from transgenic mice that express enhanced green fluorescent protein under the control of the human glial fibrillary acidic protein promoter. Astrocytes were studied by whole-cell voltage clamp. Exogenous ATP evoked inward currents in 75 of 81 astrocytes. In the majority (65%) of cells, ATP-induced responses comprising a fast and delayed component; in the remaining subpopulation of astrocytes, ATP triggered a smoother response with rapid peak and slowly decaying plateau phase. The fast component of the response was sensitive to low concentrations of ATP (with EC₅₀ of 40 nM). All ATP-induced currents were blocked by pyridoxal-phosphate-6-azophenyl-2',4'-disulfonate (PPADS); they were insensitive to ivermectin. Quantitative real-time PCR demonstrated strong expression of P2X₁ and P2X₅ receptor subunits and some expression of P2X₂ subunit mRNAs. The main properties of the ATP-induced response in cortical astrocytes (high sensitivity to ATP, biphasic kinetics, and sensitivity to PPADS) were very similar to those reported for P2X_1/5 heteromeric receptors studied previously in heterologous expression systems.

Key words: P2X_1/5 receptors; cortex; GFAP–EGFP mice; astrocytes; neuron–glial interaction; P2X

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Received Dec. 9, 2007; revised April 8, 2008; accepted April 9, 2008.

Correspondence should be addressed to Prof. Alexei Verkhratsky, Faculty of Life Sciences, The University of Manchester, 1.124 Stopford Building, Oxford Road, Manchester M13 9PT, UK. Email: alex.verkhratsky{at}manchester.ac.uk

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