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TRPA1 and TRPV4 activation in human odontoblasts stimulates ATP release.
J Dent Res. 2014 Sep; 93(9):911-7.JD

Abstract

The mechanism of pain in dentine hypersensitivity is poorly understood but proposed to result from the activation of dental sensory neurons in response to dentinal fluid movements. Odontoblasts have been suggested to contribute to thermal and mechanosensation in the tooth via expression of transient receptor potential (TRP) channels. However, a mechanism by which odontoblasts could modulate neuronal activity has not been demonstrated. In this study, we investigated functional TRP channel expression in human odontoblast-like cells and measured ATP release in response to TRP channel activation. Human immortalized dental pulp cells were driven toward an odontoblast phenotype by culture in conditioned media. Functional expression of TRP channels was determined with reverse transcription polymerase chain reaction and ratiometric calcium imaging with Fura-2. ATP release was measured using a luciferin-luciferase assay. Expression of mRNA for TRPA1, TRPV1, and TRPV4 but not TRPM8 was detected in odontoblasts by reverse transcription polymerase chain reaction. Expression of TRPV4 protein was detected by Western blotting and immunocytochemistry. The TRPA1 agonists allyl isothiocyanate and cinnamaldehyde and the TRPV4 agonist GSK1016790A caused a concentration-dependent increase in intracellular Ca(2+) concentration that was inhibited by the selective antagonists HC030031, AP18, and HC067047, respectively. In contrast, exposure to the TRPV1 agonist capsaicin or the TRPM8 agonist icilin had no effect on intracellular Ca(2+) concentration. Treatment with allyl isothiocyanate, cinnamaldehyde, or GSK1016790A caused an increase in ATP concentration in culture medium that was abolished by preincubation with TRP channel antagonists. These data demonstrate that activation of TRPA1 and TRPV4 channels in human odontoblast-like cells can stimulate ATP release. We were unable to confirm the presence of thermosensitive TRPV1 and TRPM8 that has previously been reported in odontoblasts.

Authors+Show Affiliations

Biomaterials, Tissue Engineering, and Imaging, King's College London, London, UK Department of Oral Surgery, Dental Institute, King's College London, London, UK.Wolfson Centre for Age-Related Diseases, King's College London, London, UK.Wolfson Centre for Age-Related Diseases, King's College London, London, UK.Wolfson Centre for Age-Related Diseases, King's College London, London, UK.Department of Oral Surgery, Dental Institute, King's College London, London, UK.Department of Oral Surgery, Dental Institute, King's College London, London, UK.Biomaterials, Tissue Engineering, and Imaging, King's College London, London, UK.Wolfson Centre for Age-Related Diseases, King's College London, London, UK andrew.2.grant@kcl.ac.uk.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

25062738

Citation

Egbuniwe, O, et al. "TRPA1 and TRPV4 Activation in Human Odontoblasts Stimulates ATP Release." Journal of Dental Research, vol. 93, no. 9, 2014, pp. 911-7.
Egbuniwe O, Grover S, Duggal AK, et al. TRPA1 and TRPV4 activation in human odontoblasts stimulates ATP release. J Dent Res. 2014;93(9):911-7.
Egbuniwe, O., Grover, S., Duggal, A. K., Mavroudis, A., Yazdi, M., Renton, T., Di Silvio, L., & Grant, A. D. (2014). TRPA1 and TRPV4 activation in human odontoblasts stimulates ATP release. Journal of Dental Research, 93(9), 911-7. https://doi.org/10.1177/0022034514544507
Egbuniwe O, et al. TRPA1 and TRPV4 Activation in Human Odontoblasts Stimulates ATP Release. J Dent Res. 2014;93(9):911-7. PubMed PMID: 25062738.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - TRPA1 and TRPV4 activation in human odontoblasts stimulates ATP release. AU - Egbuniwe,O, AU - Grover,S, AU - Duggal,A K, AU - Mavroudis,A, AU - Yazdi,M, AU - Renton,T, AU - Di Silvio,L, AU - Grant,A D, Y1 - 2014/07/25/ PY - 2014/7/27/entrez PY - 2014/7/27/pubmed PY - 2014/10/24/medline KW - dentin KW - ion channels KW - orofacial pain KW - sensation KW - signal transduction KW - transient receptor potential channel SP - 911 EP - 7 JF - Journal of dental research JO - J Dent Res VL - 93 IS - 9 N2 - The mechanism of pain in dentine hypersensitivity is poorly understood but proposed to result from the activation of dental sensory neurons in response to dentinal fluid movements. Odontoblasts have been suggested to contribute to thermal and mechanosensation in the tooth via expression of transient receptor potential (TRP) channels. However, a mechanism by which odontoblasts could modulate neuronal activity has not been demonstrated. In this study, we investigated functional TRP channel expression in human odontoblast-like cells and measured ATP release in response to TRP channel activation. Human immortalized dental pulp cells were driven toward an odontoblast phenotype by culture in conditioned media. Functional expression of TRP channels was determined with reverse transcription polymerase chain reaction and ratiometric calcium imaging with Fura-2. ATP release was measured using a luciferin-luciferase assay. Expression of mRNA for TRPA1, TRPV1, and TRPV4 but not TRPM8 was detected in odontoblasts by reverse transcription polymerase chain reaction. Expression of TRPV4 protein was detected by Western blotting and immunocytochemistry. The TRPA1 agonists allyl isothiocyanate and cinnamaldehyde and the TRPV4 agonist GSK1016790A caused a concentration-dependent increase in intracellular Ca(2+) concentration that was inhibited by the selective antagonists HC030031, AP18, and HC067047, respectively. In contrast, exposure to the TRPV1 agonist capsaicin or the TRPM8 agonist icilin had no effect on intracellular Ca(2+) concentration. Treatment with allyl isothiocyanate, cinnamaldehyde, or GSK1016790A caused an increase in ATP concentration in culture medium that was abolished by preincubation with TRP channel antagonists. These data demonstrate that activation of TRPA1 and TRPV4 channels in human odontoblast-like cells can stimulate ATP release. We were unable to confirm the presence of thermosensitive TRPV1 and TRPM8 that has previously been reported in odontoblasts. SN - 1544-0591 UR - https://www.unboundmedicine.com/medline/citation/25062738/TRPA1_and_TRPV4_activation_in_human_odontoblasts_stimulates_ATP_release_ L2 - https://journals.sagepub.com/doi/10.1177/0022034514544507?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -