Appearance of dentin gamma-carboxyglutamic acid-containing proteins in developing rat molars in vitro
An in vitro model of mineralization was devised in order to study the developmental appearance of dentin gamma-carboxyglutamic acid-containing proteins (DGPs) in relation to the onset of mineralization. Maxillary third molars from 11-day-old rats were cultured with or without fetal calf serum (FCS) as modified from Navia et al. Molars were incubated without radiolabel, or with either /sup 45/CaCl/sub 2/ (5 microCi/ml) for 24 hr at various stages of a ten-day culture period or (/sup 3/H)-leucine (10 microCi/ml) for 24 hr at the eighth day of culture. Molars were lyophilized and extracted with 10% formic acid overnight at 4 degrees C. DGPs in extracts were detected by immunologic and chromatographic techniques; DGPs in molar sections were detected by immunolocalization using indirect immunofluorescence. Molar development was evaluated histologically using the Von Kossa staining technique. Molars cultured with FCS showed histologic evidence for mineralized dentin and enamel and a significant increase in /sup 45/Ca uptake after the sixth day in vitro. Eleven-day-old molars in vivo and molars cultured without FCS showed no evidence of the presence of mineralized tissues. (/sup 3/H)-Leucine-labeled DGPs were isolated and identified by affinity and reversed-phase high-performance liquid chromatography and by gel electrophoresis from both mineralized and unmineralized molars. DGP antigens were localized immunohistochemically using rabbit anti-rat antibodies raised against a highly purified DGP preparation. In the unmineralized molar, antigenicity was seen in odontoblasts but not in predentin matrix, preodontoblasts, or in any other cell type. Antigens in the mineralized molar were localized to odontoblasts and dentin.
- Research Organization:
- Univ. of Alabama at Birmingham, University Station
- OSTI ID:
- 5023787
- Journal Information:
- J. Dent. Res.; (United States), Vol. 7
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
62 RADIOLOGY AND NUCLEAR MEDICINE
TEETH
MINERALIZATION
AFFINITY
ANTIBODIES
ANTIGENS
CALCIUM 45
DENTIN
DENTISTRY
ELECTROPHORESIS
ENAMELS
FLUORESCENCE
GLUTAMIC ACID
IMMUNITY
IN VITRO
LEUCINE
LIQUID COLUMN CHROMATOGRAPHY
PROTEINS
RATS
TRACER TECHNIQUES
TRITIUM COMPOUNDS
ALKALINE EARTH ISOTOPES
AMINO ACIDS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CALCIUM ISOTOPES
CARBOXYLIC ACIDS
CHROMATOGRAPHY
COATINGS
DAYS LIVING RADIOISOTOPES
DIGESTIVE SYSTEM
EVEN-ODD NUCLEI
INTERMEDIATE MASS NUCLEI
ISOTOPE APPLICATIONS
ISOTOPES
LABELLED COMPOUNDS
LUMINESCENCE
MAMMALS
MEDICINE
NUCLEI
ORAL CAVITY
ORGANIC ACIDS
ORGANIC COMPOUNDS
RADIOISOTOPES
RODENTS
SEPARATION PROCESSES
VERTEBRATES
550201* - Biochemistry- Tracer Techniques
550600 - Medicine