Odontoblasts form dentin at the outermost surface of tooth pulp. An increasing level of evidence in recent years, along with their locational advantage, implicates odontoblasts as a secondary role as sensory or immune cells. Extracellular adenosine triphosphate (ATP) is a well-characterized signaling molecule in the neuronal and immune systems, and its potential involvement in interodontoblast communications was recently demonstrated. In an effort to elaborate the ATP-mediated signaling pathway in odontoblasts, the current study performed single-cell reverse transcription polymerase chain reaction (RT-PCR) and immunofluorescent detection to investigate the expression of ATP receptors related to calcium signal in odontoblasts from incisal teeth of 8- to 10-wk-old rats, and demonstrated an in vitro response to ATP application via calcium imaging experiments. While whole tissue RT-PCR analysis detected P2Y₂, P2Y₄, and all 7 subtypes (P2X₁ to P2X₇) in tooth pulp, single-cell RT-PCR analysis of acutely isolated rat odontoblasts revealed P2Y₂, P2Y₄, P2X₂, P2X₄, P2X₆, and P2X₇ expression in only a subset (23% to 47%) of cells tested, with no evidence for P2X₁, P2X₃, and P2X₅ expression. An increase of intracellular Ca²⁺ concentration in response to 100μM ATP, which was repeated after pretreatment of thapsigargin or under the Ca²⁺-free condition, suggested function of both ionotropic and metabotropic ATP receptors in odontoblasts. The enhancement of ATP-induced calcium response by ivermectin and inhibition by 5-(3-bromophenyl)-1,3-dihydro-2H-benzofuro[3,2-e]-1,4-diazepin-2-one (5-BDBD) confirmed a functional P2X₄ subtype in odontoblasts. Positive calcium response to 2',3'-O-(benzoyl-4-benzoyl)-ATP (BzATP) and negative response to α,β-methylene ATP suggested P2X₂, P2X₄, and P2X₇ as functional subunits in rat odontoblasts. Single-cell RT-PCR analysis of the cells with confirmed calcium response and immunofluorescent detection further corroborated the expression of P2X₄ and P2X₇ in odontoblasts. Overall, this study demonstrated heterogeneous expression of calcium-related ATP receptor subtypes in subsets of individual odontoblasts, suggesting extracellular ATP as a potential signal mediator for odontoblastic functions.

#	Concept	H-Index	Publications	Citations
1	Medicine	1	1	101
2	Biochemistry	1	1	101
3	Metabolism	1	1	101
4	Clinical Chemistry	1	1	101
5	Mass Spectrometry	1	1	101

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