ADAM28 participates in the regulation of tooth development.
Disintegrin and metalloprotease (ADAM) proteins are a family of membrane-anchored glycoproteins with diverse functions in fertilisation, development, neurogenesis and protein ectodomain shedding. ADAM28 is a newly discovered member of the ADAM family in humans and murine with autocatalytic activity. Recently, the authors screened ADAM28 genes from patients with congenital hypoplasia of tooth root, and studied the relationship between ADAM28 and tooth development. A polyclonal antibody (pAb) against ADAM28 was preparared, and the expression and localisation of ADAM28 were detected in tooth germ and dental mesenchymal cells. The results indicated that the prokaryotic expression vector pGEX-4T-ADAM28 was constructed successfully. Glutathione S-transferase-ADAM28 fusion protein was generated after inducement by isopropylthio-beta-d-galactoside and isolated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The purified fusion protein was used as an antigen for production of antibody. Western blot and enzyme-linked immunosorbent assay analyses verified that the antibody had a high specificity and titre. Immunohistochemistry and reverse transcriptase-polymerase chain reaction showed that ADAM28 was expressed at each stage of tooth germ development at different levels. Moreover, it was expressed in human dental follicle cells, human dental papilla cells, human dental pulp stem cells, human periodontal ligament cells and human dental cervical loop epithelial cells at transcription level. In conclusion, it is reasonable to suggest that ADAM28 may participate in tooth development and the regulation of odontogenic mesenchymal cells through progressive reciprocal inductive interactions between the epithelium and the mesenchyme.
Zhao
Zheng
Z
Department of Paediatric Dentistry, College of Stomatology, Fourth Military Medical University, Xi'an, Shaanxi 710032, China.
Wen
Ling-Ying
LY
Jin
Ming
M
Deng
Zhi-Hong
ZH
Jin
Yan
Y
D051722:ADAM Proteins / Q000032:analysis* / Q000235:genetics; D000818:Animals; D002478:Cells, Cultured; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D005982:Glutathione Transferase / Q000235:genetics; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D008297:Male; D059630:Mesenchymal Stem Cells / Q000502:physiology; D051379:Mice; D008807:Mice, Inbred BALB C; D009805:Odontogenesis / Q000502:physiology*; D012333:RNA, Messenger / Q000235:genetics; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D014070:Tooth / Q000196:embryology*; D014083:Tooth Germ / Q000196:embryology
D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase; D051722:ADAM Proteins; C410224:ADAM28 protein, human