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Expression of P2X2 and P2X3 receptor subunits in rat carotid body afferent neurones: role in chemosensory signalling

Mona Prasad, Ian M. Fearon, Min Zhang, Michael Laing, Cathy Vollmer and Colin A. Nurse

1. Hypoxic chemotransmission in the rat carotid body (CB) is mediated in part by ATP acting on suramin-sensitive P2X purinoceptors. Here, we use RT-PCR, cloning and sequencing techniques to show P2X2 and P2X3 receptor expression in petrosal neurones, some of which develop functional chemosensory units with CB receptor clusters in co-culture.
2. Single-cell RT-PCR revealed that hypoxia-responsive neurones, identified electrophysiologically in co-culture, expressed both P2X2 and P2X3 mRNA.
3. Isohydric hypercapnia (10 % CO₂; pH 7.4) caused excitation of chemosensory units in co-culture. This excitation depended on chemical transmission, with ATP acting as a co-transmitter, since it was inhibited by reduction of the extracellular Ca²⁺:Mg²⁺ ratio and by the purinoceptor blocker suramin (50-100 µM).
4. Hypoxia and isohydric hypercapnia could separately excite the same chemosensory unit, and together the two stimuli interacted synergistically.
5. Using confocal immunofluorescence, co-localization of P2X2 and P2X3 protein was demonstrated in many petrosal somas and CB afferent terminals in situ. Taken together, these data indicate that ATP and P2X2-P2X3 purinoceptors play important roles in the peripheral control of respiration by carotid body chemoreceptors.

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