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Extracellular ATP Induces Calcium Signaling in Odontoblasts

Title
Extracellular ATP Induces Calcium Signaling in Odontoblasts
Author
정승준
Keywords
dental pulp; odontoblasts; dentin; adenosine triphosphate; single-cell analysis; purinergic receptors
Issue Date
2017-02
Publisher
SAGE PUBLICATIONS INC
Citation
JOURNAL OF DENTAL RESEARCH, v. 96, no. 2, page. 200-207
Abstract
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 P2Y(2), P2Y(4), and all 7 subtypes (P2X(1) to P2X(7)) in tooth pulp, single-cell RT-PCR analysis of acutely isolated rat odontoblasts revealed P2Y(2), P2Y(4), P2X(2), P2X(4), P2X(6), and P2X(7) expression in only a subset (23% to 47%) of cells tested, with no evidence for P2X(1), P2X(3), and P2X(5) expression. An increase of intracellular Ca2+ concentration in response to 100M 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 P2X(4) subtype in odontoblasts. Positive calcium response to 2,3-O-(benzoyl-4-benzoyl)-ATP (BzATP) and negative response to ,-methylene ATP suggested P2X(2), P2X(4), and P2X(7) 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 P2X(4) and P2X(7) 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.
URI
https://journals.sagepub.com/doi/10.1177/0022034516671308https://repository.hanyang.ac.kr/handle/20.500.11754/112915
ISSN
0022-0345; 1544-0591
DOI
10.1177/0022034516671308
Appears in Collections:
COLLEGE OF MEDICINE[S](의과대학) > MEDICINE(의학과) > Articles
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