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Modification of tooth development by heat shock protein 60

International Journal of Oral Science 2016年第1期8卷 24-31页

作者： Tamas Papp Angela Polyak Krisztina Papp Zoltan Meszar Roza Zakany Eva Meszar-Katona Paine Terdik Tiinde Chang Hwa Ham Szabolcs Felszeghy Department of Anatomy Histologyand EmbryologyFaculty of MedicineUniversity of DebrecenDebrecenHungary Scoliosis Research Institute Korea University Guro HospitalSeoul Department of Oral Anatomy Faculty of DentistryUniversity of DebrecenDebrecenHungary

several heat shock proteins have been investigated in relation to tooth development, no available information is available about the spatial and temporal expression pattern of heat shock protein 60 （Hsp 60）. To characterize Hsp 60 expression in the structures of the developing tooth germ, we used Western blotting, immunohistochemistry and in situ hybridization. Hsp 60 was present in high amounts in the inner and outer enamel epithelia, enamel knot （EK） and stratum intermedium （SI）. Hsp 60 also appeared in odontoblasts beginning in the bell stage. To obtain data on the possible effect of Hsp 60 on isolated lower incisors from mice, we performed in vitro culturing. To investigate the effect of exogenous Hsp 60 on the cell cycle during culturing, we used the 5-bromo-2- deoxyuridine （BrdU） incorporation test on dental cells. Exogenously administered Hsp 60 caused bluntness at the apical part of the 16.5-day-old tooth germs, but it did not influence the proliferation rate of dental cells. We identified the expression of Hsp 60 in the developing tooth germ, which was present in high concentrations in the inner and outer enamel epithelia, EK, SI and odontoblasts. High concentration of exogenous Hsp 60 can cause abnormal morphology of the tooth germ, but it did not influence the proliferation rate of the dental cells. Our results suggest that increased levels of Hsp 60 may cause abnormalities in the morphological development of the tooth germ and support the data on the significance of Hsp during the developmental processes.

关键词： enamel organ heat shock protein 60 inhibitor of κB kinase morphology mouse tooth development

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Single cell RNA sequencing reveals mesenchymal heterogeneity and critical functions of Cd271 in tooth development

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World Journal of Stem Cells 2023年第6期15卷 589-605页

作者： Yan-Yan Zhang Feng Li Xiao-Ke Zeng Yan-Hui Zou Bing-Bing Zhu Jia-Jia Ye Yun-Xiao Zhang Qiu Jin Xin Nie School&Hospital of Stomatology Wenzhou Medical UniversityWenzhou 325027Zhejiang ProvinceChina

BACKGROUND Accumulating evidence suggests that the maxillary process,to which cranial crest cells migrate,is essential to tooth *** studies indicate that Cd271 plays an essential role in ***,the underlying mechanisms have yet to be *** To establish the functionally heterogeneous population in the maxillary process,elucidate the effects of Cd271 deficiency on gene expression *** p75NTR knockout(Cd271-/-)mice(from American Jackson laboratory)were used to collect the maxillofacial process tissue of p75NTR knockout mice,and the wildtype maxillofacial process of the same pregnant mouse wild was used as *** single cell suspension,the cDNA was prepared by loading the single cell suspension into the 10x Genomics Chromium system to be sequenced by NovaSeq6000 sequencing ***,the sequencing data in Fastq format were *** FastQC software is used to evaluate the quality of data and CellRanger analyzed the *** gene expression matrix is read by R software,and Seurat is used to control and standardize the data,reduce the dimension and *** search for marker genes for subgroup annotation by consulting literature and database;explore the effect of p75NTR knockout on mesenchymal stem cells(MSCs)gene expression and cell proportion by cell subgrouping,differential gene analysis,enrichment analysis and protein-protein interaction network analysis;understand the interaction between MSCs cells and the differentiation trajectory and gene change characteristics of p75NTR knockout MSCs by cell communication analysis and pseudo-time *** we verified the findings single cell sequencing in *** We identified 21 cell clusters,and we re-clustered these into three ***,we revealed the cell–cell communication networks between *** clarified that Cd271 was significantly associated with the regulation of *** This study provides comprehe

关键词： Cd271 Mesenchymal stem cells Single cell RNA sequencing Osteogenesis Mineralization tooth development

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Phenotypic analyses of Noggin mutant teeth and gene expression patternssuggest a functional redundancy among BMP antagonists during tooth development

Phenotypic analyses of Noggin mutant teeth and gene expressi...

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“细胞活动生命活力”——中国细胞生物学学会全体会员代表大会暨第十二次学术大会

作者： Xuefeng Hu~1,Ying Wang~(2,3),Fenglei He~3,Lu Li~3,Yanding Zhang~1,YiPing Chen~(1,3) 1.Fujian Key Laboratory of developmental and Neuro Biology,College of Life Sciences,Fujian Normal University Fuzhou,Fujian Province,P.R.China 2.College of Stomatology,The Fourth Military Medical University,Xi’an,Shaangxi,P.R.China 3.Department of Cell and Molecular Biology,Tulane University New Orleans,LA 70118,USA

BMP signaling plays crucial roles in the development of many organs,including the *** important is BMP signaling homeostasis,as demonstrated by a large spectrum of organdefects in mice lacking the extracellular BMP antagonist ***,we examine Noggin expression pattern in the developing teeth and analyze tooth phenotype in Noggin mutant *** is initially expressed in the maxillary mesenchyme adjunct to the upper incisor at theinitiation stage,and then in the developing teeth including incisors and molars from the bud *** mutant mice develop normal molars and mandibular incisors,but form a single,medially located upper incisor that is arrested at the late bud stage. We further show that Chordin and Gremlin,the other two BMP antagonists,exhibit an overlapping expression pattern with that of Noggin in the developing *** observations suggest a functional redundancy between these BMP antagonists during tooth development

关键词： BMP antagonist Noggin Chordin Gremlin tooth development

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Molecular regulatory mechanism of tooth root development

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International Journal of Oral Science 2012年第4期4卷 177-181页

作者： Xiao-Feng Huang Yang Chai Department of Stomatology Beijing Friendship HospitalCapital Medical University Center for Craniofacial Molecular Biology Ostrow School of DentistryUniversity of Southern California

The root is crucial for the physiological function of the tooth, and a healthy root allows an artificial crown to function as required clinically. tooth crown development has been studied intensively during the last few decades, but root development remains not well understood. Here we review the root development processes, including cell fate determination, induction of odontoblast and cementoblast differentiation, interaction of root epithelium and mesenchyme, and other molecular mechanisms. This review summarizes our current understanding of the signaling cascades and mechanisms involved in root development. It also sets the stage for de novo tooth regeneration.

关键词： Hertwig's epithelial root sheath periodontium root development tooth development

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Globoside accelerates the differentiation of dental epithelial cells into ameloblasts

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International Journal of Oral Science 2016年第4期8卷 205-212页

作者： Takashi Nakamura Yuta Chiba Masahiro Naruse Kan Saito Hidemitsu Harada Satoshi Fukumoto Division of Molecular Pharmacology and Cell Biophysics Department of Oral Biology Tohoku University Graduate School of Dentistry Division of Pediatric Dentistry Department of Oral Health and Development Sciences Tohoku University Graduate School of Dentistry Division of developmental Biology and Regenerative Medicine Department of Anatomy Iwate Medical University

tooth crown morphogenesis is tightly regulated by the proliferation and differentiation of dental epithelial cells. Globoside （Gb4）, a globo-series glycosphingolipid, is highly expressed during embryogenesis as well as organogenesis, including tooth development. We previously reported that Gb4 is dominantly expressed in the neutral lipid fraction of dental epithelial cells. However, because its functional role in tooth development remains unknown, we investigated the involvement of Gb4 in dental epithelial cell differentiation. The expression of Gb4 was detected in ameloblasts of postnatal mouse molars and incisors. A cell culture analysis using HAT-7 cells, a rat-derived dental epithelial cell line, revealed that Gb4 did not promote dental epithelial cell proliferation. Interestingly, exogenous administration of Gb4 enhanced the gene expression of enamel extracellular matrix proteins such as ameloblastin, amelogenin, and enamelin in dental epithelial cells as well as in developing tooth germs. Gb4 also induced the expression of TrkB, one of the key receptors required for ameloblast induction in dental epithelial cells. In contrast, Gb4 downregulated the expression of p75, a receptor for neurotrophins （including neurotrophin-4） and a marker of undifferentiated dental epithelial cells. In addition, we found that exogenous administration of Gb4 to dental epithelial cells stimulated the extracellular signal-regulated kinase and p38 mitogen-activated protein kinase signalling pathways. Furthermore, Gb4 induced the expression of epiprofin and Runx2, the positive regulators for ameloblastin gene transcription. Thus, our results suggest that Gb4 contributes to promoting the differentiation of dental epithelial cells into ameloblasts.

关键词： ameloblast differentiation enamel matrix epiprofin glycosphingolipids tooth development

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Spatial signalling mediated by the transforming growth factor-β signalling pathway during tooth formation

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International Journal of Oral Science 2016年第4期8卷 199-204页

作者： Xin-Yu He Ke Sun Ruo-Shi Xu Jia-Li Tan Cai-Xia Pi Mian Wan Yi-Ran Peng Ling Ye Li-Wei Zheng Xue-Dong Zhou State Key Laboratory of Oral Diseases West China Hospital of Stomatology Sichuan University West China School of Stomatology Sichuan University Guanghua School of Stomatology Hospital of Stomatology Department of Endodontics Guangdong Provincial Key Laboratory of Stomatology Sun Yat-sen University

tooth development relies on sequential and reciprocal interactions between the epithelial and mesenchymal tissues, and it is continuously regulated by a variety of conserved and specific temporal-spatial signalling pathways. It is well known that suspensions of tooth germ cells can form tooth-like structures after losing the positional information provided by the epithelial and mesenchymal tissues. However, the particular stage in which the tooth germ cells start to form tooth-like structures after losing their positional information remains unclear. In this study, we investigated the reassociation of tooth germ cells suspension from different morphological stages during tooth development and the phosphorylation of Smad2/3 in this process. Four tooth morphological stages were designed in this study. The results showed that tooth germ cells formed odontogenic tissue at embryonic day （E） 14.5, which is referred to as the cap stage, and they formed tooth-like structures at E16.5, which is referred to as the early bell stage, and E18.5, which is referred to as the late bell stage. Moreover, the transforming growth factor-β signalling pathway might play a role in this process.

关键词： positional information transforming growth factor-13 signalling pathway tooth development

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Methods for studying tooth root cementum by light microscopy

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International Journal of Oral Science 2012年第3期4卷 119-128页

作者： Brian L Foster National Institute for Arthritis and Musculoskeletal and Skin Diseases (NIAMS) National Institutes of Health (NIH)

The tooth root cementum is a thin, mineralized tissue covering the root dentin that is present primarily as acellular cementum on the cervical root and cellular cementum covering the apical root. While cementum shares many properties in common with bone and dentin, it is a unique mineralized tissue and acellular cementum is critical for attachment of the tooth to the surrounding periodontal ligament （PDL）. Resources for methodologies for hard tissues often overlook cementum and approaches that may be of value for studying this tissue. To address this issue, this report offers detailed methodology, as well as comparisons of several histological and immunohistochemical stains available for imaging the cementum-PDL complex by light microscopy. Notably, the infrequently used Alcian blue stain with nuclear fast red counterstain provided utility in imaging cementum in mouse, porcine and human teeth. While no truly unique extracellular matrix markers have been identified to differentiate cementum from the other hard tissues, immunohistochemistry for detection of bone sialoprotein （BSP）, osteopontin （OPN）, and dentin matrix protein 1 （DMP1） is a reliable approach for studying both acellular and cellular cementum and providing insight into developmental biology of these tissues. Histoloeical and immunohistochemical aooroaches Drovide insight on developmental biology of cementum.

关键词： bone bone sialoprotein cementum dentin dentin matrix protein 1 osteopontin periodontal ligament tooth development

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Making a tooth:growth factors,transcription factors,and stem cells

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Cell Research 2005年第5期15卷 301-316页

作者： Yan Ding ZHANG1,2, Zhi CHEN1, Yi Qiang SONG3, Chao LIU2, Yi Ping CHEN2,3,* 1College of Stomatology, Wuhan University, Wuhan 430072, China 2College of Bioengineering, Fujian Normal University, Fuzhou 350007, China 3Division of developmental Biology, Department of Cell and Molecular Biology, Tulane University, New Orleans, LA 70118, USA College of Stomatology Wuhan University Wuhan China College of Bioengineering Fujian Normal University Fuzhou China Division of developmental Biology Department of Cell and Molecular Biology Tulane University New Orleans USA

Mammalian tooth development is largely dependent on sequential and reciprocal epithelial-mesenchymal interactions. These processes involve a series of inductive and permissive interactions that result in the determination, differentiation, and organization of odontogenic tissues. Multiple signaling molecules, including BMPs, FGFs, Shh, and Wnt proteins, have been implicated in mediating these tissue interactions. Transcription factors participate in epithelial-mesenchymal interactions via linking the signaling loops between tissue layers by responding to inductive signals and regulating the expression of other signaling molecules. Adult stem cells are highly plastic and multipotent. These cells including dental pulp stem cells and bone marrow stromal cells could be reprogrammed into odontogenic fate and participated in tooth formation. Recent progress in the studies of molecular basis of tooth development, adult stem cell biology, and regene- ration will provide fundamental knowledge for the realization of human tooth regeneration in the near future.

关键词： tooth development transcription factor growth factor stem cells regeneration.

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Klf4 Promote Odontoblasts Differentiation By Targeting Dmp1

Klf4 Promote Odontoblasts Differentiation By Targeting Dmp1

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第二届全国发育生物学大会

作者： Heng Lin Zheyi Sunday Lu Zhang Zhi Chen School and Hospital of Stomatology Wuhan University

Kruppel-like factor 4(Klf4)is a member of Klf zink finger transcriptional factor *** Klf4 plays a pivotal role in ***,we have found that Klf4 was expressed in the polarizing and elongating *** the function of Klf4 in the differentiation of odontoblasts and osteoblasts is still *** hypothesized Klf4 promoted the differentiation of odontoblasts by up-regulating some odontoblast-related *** containing inactivated Klf4 in their neural crest cells(Wnt1-Cre;Klf4)were obtained for phenotype *** head samples were dissected for histological and immunostaining or for X-ray *** and loss of function of Klf4was applied in mDPC6T cell ***,ChIP and DLA assays were used to detect the target gene of *** showed that,for the conditional knockout mice Dmpl and Dsp were downregulated in odontoblasts of PN0.5 molar germ.H-E staining showed thicken predentin layer and sparse alveolar bone in the conditional knockout *** conditional knockout mice also showed weaker blue in the dentin and alveolar bone for the azan *** high resolution X-ray,we have found decrease of the grey scale in the dentin but not the enamel in the conditional knockout *** vitro overexpression of Klf4 in mDPC6T and mc-3T3-E1significantly up-regulated Dmpl,Dspp,and ***-down of Klf4 down-regulated expression of Dmp1,Dspp,and *** on analysis of ChIP data,EMSA and dual luciferase activity assays,we confirmed that Klf4 was able to specifically bind to the Dmpl promoter and transactivate its ***4 is essential in the odontoblasts differentiation and dentin mineralization.

关键词： Klf4 odontoblast tooth development dentin mineralization.

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