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J Biol Chem, Vol. 273, Issue 41, 26799-26808, October 9, 1998

Expression and Trans-synaptic Regulation of P_2x4 and P_2z Receptors for Extracellular ATP in Parotid Acinar Cells
EFFECTS OF PARASYMPATHETIC DENERVATION

From the Department of Neuroscience, Tufts University Medical School, Boston, Massachusetts 02111

Trans-synaptic regulation of muscarinic, peptidergic, and purinergic responses after denervation has been reported previously in rat parotid acinar cells (McMillian, M. K., Soltoff, S. P., Cantley, L. C., Rudel, R., and Talamo, B. R. (1993) Br. J. Pharmacol. 108, 453-461). Characteristics of the ATP-mediated responses and the effects of parasympathetic denervation were further analyzed through assay of Ca²⁺ influx, using fluorescence ratio imaging methods, and by analysis of P_2x receptor expression. ATP activates both a high affinity and a low affinity response with properties corresponding to the recently described P_2x4 and the P_2z (P_2x7)-type purinoceptors, respectively. Reverse transcription-polymerase chain reaction analysis reveals mRNA for P_2x4 as well as P_2x7 subtypes but not P_2x1, P_2x2, P_2x3, P_2x5, or P_2x6. P_2x4 protein also is detected by Western blotting. Distribution of the two types of ATP receptor responses on individual cells was stochastic, with both high and low affinity responses on some cells, and only a single type of response on others. Sensitivity to P_2x4-type activation also varied even among cells responsive to low concentrations of ATP. Parasympathetic denervation greatly enhanced responses, tripling the proportion of acinar cells with a P_2x4-type response and increasing the fraction of highly sensitive cells by 7-fold. Moreover, P_2x4 mRNA is significantly increased following parasympathetic denervation. These data indicate that sensitivity to ATP is modulated by neurotransmission at parasympathetic synapses, at least in part through increased expression of P_2x4 mRNA, and suggest that similar regulation may occur at other sites in the nervous system where P_2x4 receptors are widely expressed.

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