Immunoblot detection and expression of enamel proteins at the apical portion of the forming root in porcine permanent incisor tooth germs.
There have been many immunohistochemical studies of enamel proteins during root formation. In the present article, the detection and expression of enamel proteins in tissue samples prepared from the apical portion of the forming root (APFR) in porcine permanent incisor tooth germs were studied. Amelogenin, enamelin, and sheathlin were detected by immunoblot analysis, but only in small amounts. The detection of their derivatives indicated their degradation. It is, at present, unclear as to which proteinases are involved in these degradations, because activity of enamel matrix serine proteinase 1 and enamelysin was not detected on gelatin and casein zymograms. The expression of enamel proteins was also proved in the APFR sample by the detection of polymerase chain reaction products of their cDNAs, and this may be related to cells of fragmentized Hertwig's epithelial root sheath. Amelogenin expression was not greater than that of enamelin and sheathlin. It was different from the expression pattern of secretory ameloblasts involved in enamel matrix formation. These results suggest that the amelogenins found in the APFR do not form a three-dimensional structure of amelogenin micelles, which has been proposed for the secretory enamel matrix structure. In this case, the enamel proteins could spread out easily following degradation into the matrix of future cementum. Some of their derivatives may play a role in the formation of the cementum.
Fukae
M
M
Department of Biochemistry, School of Dental Medicine, Tsurumi University, Yokohama, Japan.
Tanabe
T
T
Yamakoshi
Y
Y
Yamada
M
M
Ujiie
Y
Y
Oida
S
S
D053523:Amelogenin; D000818:Animals; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D003743:Dental Enamel / Q000254:growth & development / Q000378:metabolism; D003746:Dental Enamel Proteins / Q000235:genetics* / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D015151:Immunoblotting; D007180:Incisor / Q000254:growth & development / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013552:Swine; D019227:Tooth Apex / Q000254:growth & development* / Q000378:metabolism*; D014083:Tooth Germ / Q000254:growth & development / Q000378:metabolism*
C490889:AMBN protein, human; D053523:Amelogenin; D017931:DNA Primers; D003746:Dental Enamel Proteins; D012333:RNA, Messenger; C490891:ameloblastin protein, Sus scrofa; C038456:tuftelin