Specific amelogenin gene splice products have signaling effects on cells in culture and in implants in vivo.
Low molecular mass amelogenin-related polypeptides extracted from mineralized dentin have the ability to affect the differentiation pathway of embryonic muscle fibroblasts in culture and lead to the formation of mineralized matrix in in vivo implants. The objective of the present study was to determine whether the bioactive peptides could have been amelogenin protein degradation products or specific amelogenin gene splice products. Thus, the splice products were prepared, and their activities were determined in vitro and in vivo. A rat incisor tooth odontoblast pulp cDNA library was screened using probes based on the peptide amino acid sequencing data. Two specific cDNAs comprised from amelogenin gene exons 2,3,4,5,6d,7 and 2,3,5,6d, 7 were identified. The corresponding recombinant proteins, designated r[A+4] (8.1 kDa) and r[A-4] (6.9 kDa), were produced. Both peptides enhanced in vitro sulfate incorporation into proteoglycan, the induction of type II collagen, and Sox9 or Cbfa1 mRNA expression. In vivo implant assays demonstrated implant mineralization accompanied by vascularization and the presence of the bone matrix proteins, BSP and BAG-75. We postulate that during tooth development these specific amelogenin gene splice products, [A+4] and [A-4], may have a role in preodontoblast maturation. The [A+4] and [A-4] may thus be tissue-specific epithelial mesenchymal signaling molecules.
Veis
A
A
Department of Basic and Behavioral Sciences, Northwestern University Dental School, Chicago, Illinois 60611, USA. aveis@northwestern.edu
Tompkins
K
K
Alvares
K
K
Wei
K
K
Wang
L
L
Wang
X S
XS
Brownell
A G
AG
Jengh
S M
SM
Healy
K E
KE
D053523:Amelogenin; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D003094:Collagen / Q000235:genetics; D003746:Dental Enamel Proteins / Q000235:genetics / Q000502:physiology*; D003757:Dental Implantation; D008969:Molecular Sequence Data; D009805:Odontogenesis; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D020318:Rats, Long-Evans; D011994:Recombinant Proteins / Q000494:pharmacology
D053523:Amelogenin; C507201:Amelx protein, rat; D003746:Dental Enamel Proteins; D012333:RNA, Messenger; D011994:Recombinant Proteins; D003094:Collagen