Amelogenesis in vitro: a model for studies of epithelial postsecretory processing during tissue-specific extracellular matrix biomineralization.
The extracellular matrix (ECM) of developing mammalian enamel comprises a complex of unusual epithelial-derived proteins, which appear to function in concert to initiate and propagate tissue-specific biomineralization. Following enamel protein synthesis by ameloblast cells within the enamel organ, the subsequent steps of posttranslational modification, secretion, postsecretory processing and eventual removal of these proteins from forming enamel are largely unknown. To address this issue we have designed studies to investigate the hypothesis that enamel proteins are removed from enamel and translocated into the vasculature as relatively high-molecular-weight components. We examined enamel proteins recovered from serumless medium during prolonged organ culture of mouse capstage mandibular first molars. By 21 days in vitro the tooth crown formed and dentine and enamel biomineralization were apparent. At 31 days, explants retained metabolic activity and the enamel matrix showed extensive transformation. Immunologically identified enamel proteins of 26-18 k Da were produced by cultured tooth organs, translocated from tooth explants to the culture medium, recovered from the medium and then compared to control enamel protein from in vivo preparations. Comparable postsecretory processing of the 26-k Da amelogenin protein was observed in vitro and in vivo. We speculate that the pathway reported in the present studies is comparable to the processing of the enamel protein polypeptides of the maturing enamel which occurs in vivo. The in vitro organ culture model described in this report provides an approach with which to investigate the molecular events associated with epithelial-derived postsecretory processing of ECM molecules associated with tissue-specific biomineralization.
Fincham
A G
AG
Department of Basic Science, School of Dentistry, University of Southern California, Los Angeles 90098-0191.
Bessem
C C
CC
Bringas
P
P
Hu
Y Y
YY
Snead
M L
ML
Slavkin
H C
HC
D000565:Ameloblasts / Q000166:cytology / Q000378:metabolism*; D000566:Amelogenesis; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002478:Cells, Cultured; D003470:Culture Media / Q000032:analysis; D003743:Dental Enamel / Q000032:analysis / Q000378:metabolism; D004847:Epithelial Cells; D004848:Epithelium / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism*; D005260:Female; D007526:Isoelectric Point; D051379:Mice; D008815:Mice, Inbred Strains; D008903:Minerals / Q000378:metabolism*; D011247:Pregnancy; D011506:Proteins / Q000032:analysis
D000596:Amino Acids; D003470:Culture Media; D008903:Minerals; D011506:Proteins