Appearance of dentin gamma-carboxyglutamic acid-containing proteins in developing rat molars in vitro

Journal Article · Mon Jul 01 04:00:00 EDT 1985 · J. Dent. Res.; (United States)

OSTI ID:5023787

Finkelman, R D; Butler, W T

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), Journal Name: J. Dent. Res.; (United States) Vol. 7; ISSN JDREA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
550600 -- Medicine
59 BASIC BIOLOGICAL SCIENCES
62 RADIOLOGY AND NUCLEAR MEDICINE
AFFINITY
ALKALINE EARTH ISOTOPES
AMINO ACIDS
ANIMALS
ANTIBODIES
ANTIGENS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CALCIUM 45
CALCIUM ISOTOPES
CARBOXYLIC ACIDS
CHROMATOGRAPHY
COATINGS
DAYS LIVING RADIOISOTOPES
DENTIN
DENTISTRY
DIGESTIVE SYSTEM
ELECTROPHORESIS
ENAMELS
EVEN-ODD NUCLEI
FLUORESCENCE
GLUTAMIC ACID
IMMUNITY
IN VITRO
INTERMEDIATE MASS NUCLEI
ISOTOPE APPLICATIONS
ISOTOPES
LABELLED COMPOUNDS
LEUCINE
LIQUID COLUMN CHROMATOGRAPHY
LUMINESCENCE
MAMMALS
MEDICINE
MINERALIZATION
NUCLEI
ORAL CAVITY
ORGANIC ACIDS
ORGANIC COMPOUNDS
PROTEINS
RADIOISOTOPES
RATS
RODENTS
SEPARATION PROCESSES
TEETH
TRACER TECHNIQUES
TRITIUM COMPOUNDS
VERTEBRATES