Expression of Pannexin3 in human odontoblast-like cells and its hemichannel function in mediating ATP release

• Pannexin3 mRNA and protein are detected in the human dental pulp tissue and hOBs.
• To examine the function of Panx3, Panx3 over expression and knockdown hOBs were established.
• In presence of cold stimulus, ATP was released from hOBs into extracellular space.
• Panx3 hemichannel is one of the major ATP release channels in hOBs.

ObjectiveThe aim of this study is to investigate the expression of pannexin3 (Panx3) in human odontoblast-like cells (hOBs) and its hemichannel function in mediating ATP release.MethodsRT-PCR and immunofluorescence analysis were used to detect the expression of pannexins (Panxs) in human dental pulp tissue and cultured cells. To determine the role of Panx3 in ATP release, hOBs were infected with Panx3-overexpression lentivirus, Panx3-shRNA lentivirus or control lentivirus and then stimulated with cold buffer. Intracellular ATP was monitored using quinacrine, and then semi-quantitatively analyzed. In the meantime, the ATP release was quantitatively analyzed using the bioluminescence method when the cells were exposed to cold stimulus.ResultsPanx3 mRNA and protein were found in dental pulp tissue and cultured cells. Upon cold stimulus, intracellular ATP was released into the extracellular space. Overexpression of Panx3 accelerated ATP release, whereas inhibition of Panx3 suppressed this process.ConclusionPanx3 hemichannel is expressed in human odontoblast-like cells and mediates ATP release into the extracellular space.