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


METHODS
RT-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.


RESULTS
Panx3 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.


CONCLUSIONS
Panx3 hemichannel is expressed in human odontoblast-like cells and mediates ATP release into the extracellular space.
The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) & Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology, Wuhan University, Wuhan, Hubei, People's Republic of China; Department of Stomatology, Renmin Hospital of Wuhan University, 238#, Jiefang Road, Wuhan, Hubei, People's Republic of China.
Dongjie
D
Fu
The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) & Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology, Wuhan University, Wuhan, Hubei, People's Republic of China.
Fangfang
F
Song
The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) & Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology, Wuhan University, Wuhan, Hubei, People's Republic of China.
Hualing
H
Sun
Department of Prosthodontics, Stomatological Hospital, College of Medicine, Xi'an Jiaotong University, Xi'an, Shaanxi, People's Republic of China.
Dandan
D
Pei
The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) & Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology, Wuhan University, Wuhan, Hubei, People's Republic of China.
Yake
Y
Wang
The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) & Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology, Wuhan University, Wuhan, Hubei, People's Republic of China.
Jinmei
J
Lei
The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) & Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology, Wuhan University, Wuhan, Hubei, People's Republic of China. Electronic address: huangcui@whu.edu.cn.
Cui
C
Huang
D000255:Adenosine Triphosphate / Q000378:metabolism*; D000293:Adolescent; D000328:Adult; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D002478:Cells, Cultured; D003080:Cold Temperature; D017630:Connexins / Q000096:biosynthesis* / Q000235:genetics; D003782:Dental Pulp / Q000166:cytology / Q000378:metabolism; D003807:Dentin Sensitivity / Q000235:genetics / Q000378:metabolism; D055785:Gene Knockdown Techniques; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D009804:Odontoblasts / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000008:administration & dosage / Q000235:genetics; D055815:Young Adult
D017630:Connexins; C492766:PANX3 protein, human; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D000255:Adenosine Triphosphate
ATP release; Dentin hypersensitivity; Hemichannel; Human odontoblast-like cell (hOB); Pannexin3 (Panx3)