Knocking out P2X receptors reduces transmitter secretion in taste buds

Yijen A. Huang, Leslie M. Stone, Elizabeth Pereira, Ruibiao Yang, John C. Kinnamon, Gennady Dvoryanchikov, Nirupa Chaudhari, Thomas E. Finger, Sue C. Kinnamon, Stephen D Roper

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

In response to gustatory stimulation, taste bud cells release a transmitter, ATP, that activates P2X2 and P2X3 receptors on gustatory afferent fibers. Taste behavior and gustatory neural responses are largely abolished in mice lacking P2X2 and P2X3 receptors [P2X2 and P2X3 double knock-out (DKO) mice]. The assumption has been that eliminating P2X2 and P2X3 receptors only removes postsynaptic targets but that transmitter secretion in mice is normal. Using functional imaging, ATP biosensor cells, and a cell-free assay for ATP, we tested this assumption. Surprisingly, although gustatory stimulation mobilizes Ca 2+ in taste Receptor (Type II) cells from DKO mice, as from wild-type (WT) mice, taste cells from DKO mice fail to release ATP when stimulated with tastants. ATP release could be elicited by depolarizing DKO Receptor cells with KCl, suggesting that ATP-release machinery remains functional in DKO taste buds. To explore the difference in ATP release across genotypes, we used reverse transcriptase (RT)-PCR, immunostaining, and histochemistry for key proteins underlying ATP secretion and degradation: Pannexin1, TRPM5, and NTPDase2 (ecto-ATPase) are indistinguishable between WTandDKOmice. The ultrastructure of contacts between taste cells and nerve fibers is also normal in theDKOmice. Finally, quantitative RT-PCR show that P2X4 and P2X7, potential modulators of ATP secretion, are similarly expressed in taste buds in WT and DKO taste buds. Importantly, we find that P2X2 is expressed in WT taste buds and appears to function as an autocrine, positive feedback signal to amplify taste-evoked ATP secretion.

Original languageEnglish
Pages (from-to)13654-13661
Number of pages8
JournalJournal of Neuroscience
Volume31
Issue number38
DOIs
StatePublished - Sep 21 2011

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Taste Buds
Adenosine Triphosphate
Purinergic P2X2 Receptors
Purinergic P2X3 Receptors
Knockout Mice
Reverse Transcriptase Polymerase Chain Reaction
Biosensing Techniques
Nerve Fibers
Genotype

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Huang, Y. A., Stone, L. M., Pereira, E., Yang, R., Kinnamon, J. C., Dvoryanchikov, G., ... Roper, S. D. (2011). Knocking out P2X receptors reduces transmitter secretion in taste buds. Journal of Neuroscience, 31(38), 13654-13661. https://doi.org/10.1523/JNEUROSCI.3356-11.2011

Knocking out P2X receptors reduces transmitter secretion in taste buds. / Huang, Yijen A.; Stone, Leslie M.; Pereira, Elizabeth; Yang, Ruibiao; Kinnamon, John C.; Dvoryanchikov, Gennady; Chaudhari, Nirupa; Finger, Thomas E.; Kinnamon, Sue C.; Roper, Stephen D.

In: Journal of Neuroscience, Vol. 31, No. 38, 21.09.2011, p. 13654-13661.

Research output: Contribution to journalArticle

Huang, YA, Stone, LM, Pereira, E, Yang, R, Kinnamon, JC, Dvoryanchikov, G, Chaudhari, N, Finger, TE, Kinnamon, SC & Roper, SD 2011, 'Knocking out P2X receptors reduces transmitter secretion in taste buds', Journal of Neuroscience, vol. 31, no. 38, pp. 13654-13661. https://doi.org/10.1523/JNEUROSCI.3356-11.2011
Huang YA, Stone LM, Pereira E, Yang R, Kinnamon JC, Dvoryanchikov G et al. Knocking out P2X receptors reduces transmitter secretion in taste buds. Journal of Neuroscience. 2011 Sep 21;31(38):13654-13661. https://doi.org/10.1523/JNEUROSCI.3356-11.2011
Huang, Yijen A. ; Stone, Leslie M. ; Pereira, Elizabeth ; Yang, Ruibiao ; Kinnamon, John C. ; Dvoryanchikov, Gennady ; Chaudhari, Nirupa ; Finger, Thomas E. ; Kinnamon, Sue C. ; Roper, Stephen D. / Knocking out P2X receptors reduces transmitter secretion in taste buds. In: Journal of Neuroscience. 2011 ; Vol. 31, No. 38. pp. 13654-13661.
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