title: Transcriptional factor ATF6 is involved in odontoblastic differentiation.
abstract: ATF6 is an endoplasmic reticulum (ER) membrane-bound transcription factor that regulates various cellular functions. The purpose of this study was to investigate the role of ATF6 in odontoblast differentiation. Rat tooth germs were isolated, changes in gene expression were evaluated over time, and localization of ATF6 was determined by immunohistochemistry. Human dental pulp cells (HDPCs) were cultured with 50 µg/mL ascorbic acid and 5 mmol/L β-glycerophosphate or 100 ng/mL bone morphogenetic protein 2 to induce differentiation. Translocation of ATF6 was observed by immunofluorescence and confocal microscopy. Overexpression of ATF6 was performed with an adenoviral vector. Matrix mineralization was evaluated by alizarin red staining. Immunoreactivity to anti-ATF6 was observed in the odontoblastic layer of the molar tooth germ, and expressions of ATF6, dentin sialophosphoprotein (DSPP) and dentin matrix protein 1 (DMP1) increased gradually during tooth germ development. When HDPCs were cultured in differentiation media, ATF6, DSPP, and DMP1 expression increased with the expression of unfolded protein response (UPR) markers, BiP and CHOP. Immunofluorescence results showed that ATF6 protein moved from cytoplasm to nucleus when cells were exposed to differentiation media. Notably, overexpression of ATF6 increased DSPP and DMP1 expression, alkaline phosphatase (ALP) activity, and matrix mineralization in HDPC cultures. Inhibition of ATF6 decreased ALP activity and mineralization. These results suggest that ER membrane-bound transcriptional factor ATF6 may be involved in odontoblastic differentiation.
authors.affiliation: Department of Conservative Dentistry, School of Dentistry, Dental Science Research Institute, Chonnam National University, Gwangju, Korea.
authors.forename: J W
authors.initials: JW
authors.lastname: Kim
authors.forename: H
authors.initials: H
authors.lastname: Choi
authors.forename: B C
authors.initials: BC
authors.lastname: Jeong
authors.forename: S H
authors.initials: SH
authors.lastname: Oh
authors.forename: S W
authors.initials: SW
authors.lastname: Hur
authors.forename: B N
authors.initials: BN
authors.lastname: Lee
authors.forename: S H
authors.initials: SH
authors.lastname: Kim
authors.forename: J E
authors.initials: JE
authors.lastname: Nör
authors.forename: J T
authors.initials: JT
authors.lastname: Koh
authors.forename: Y C
authors.initials: YC
authors.lastname: Hwang
mesh_terms: D051702:Activating Transcription Factor 6 / Q000032:analysis / Q000502:physiology*; D000256:Adenoviridae / Q000235:genetics; D000469:Alkaline Phosphatase / Q000032:analysis; D000818:Animals; D055396:Bone Morphogenetic Protein 2 / Q000494:pharmacology; D002113:Calcification, Physiologic / Q000502:physiology; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D002460:Cell Line; D002467:Cell Nucleus / Q000648:ultrastructure; D003593:Cytoplasm / Q000648:ultrastructure; D003782:Dental Pulp / Q000166:cytology; D016326:Extracellular Matrix Proteins / Q000032:analysis; D005786:Gene Expression Regulation / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D009804:Odontoblasts / Q000187:drug effects / Q000502:physiology*; D010750:Phosphoproteins / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D012795:Sialoglycoproteins / Q000032:analysis; D014083:Tooth Germ / Q000166:cytology / Q000254:growth & development; D051742:Transcription Factor CHOP / Q000032:analysis; D056811:Unfolded Protein Response / Q000502:physiology; D000099311:Dentin Sialophosphoprotein
chemical_list: D051702:Activating Transcription Factor 6; D000469:Alkaline Phosphatase; D055396:Bone Morphogenetic Protein 2; D016326:Extracellular Matrix Proteins; D010750:Phosphoproteins; D012795:Sialoglycoproteins; D051742:Transcription Factor CHOP; D000099311:Dentin Sialophosphoprotein; C497111:ATF6 protein, human; C497113:Atf6 protein, rat; C525718:BMP2 protein, human; C497249:DDIT3 protein, human; C513309:DMP1 protein, human; C081635:Dmp1 protein, rat
keywords: DMP1; DSPP; ER stress; matrix mineralization; tooth development; unfolded protein response