The effects of ascorbic acid deficiency on collagen synthesis by mouse molar tooth germs in organ culture

Development ◽  
1977 ◽  
Vol 37 (1) ◽  
pp. 49-57
Author(s):  
John R. Schiltz ◽  
Joel Rosenbloom ◽  
Gordon E. Levenson
Keyword(s):  
Ascorbic Acid ◽  
Organ Culture ◽  
Culture Medium ◽  
Large Fraction ◽  
Culture Conditions ◽  
Molar Tooth ◽  
Second Molar ◽  
Acid Deficiency ◽  
Tooth Germs

Second molar tooth germs from 2-day-old Swiss-Webster mice, grown in organ culture for 7 days in ascorbic-acid-deficient medium, synthesized about 65 % as much protein (measured by incorporation of [14C]proline during a 24-h pulse) as did ascorbic-acid-supplemented controls. The newly synthesized proteins from ascorbic-acid-deficient cultures contained only about 7% of the hydroxyproline content of the controls. Collagenase digestion of the newly synthesized proteins showed that collagen comprised the same fraction of the total protein synthesized under both culture conditions. This result indicates that the ascorbatedeficient cultures made significant quantities of underhydroxylated collagen. Partial characterization of the collagen alpha chains on carboxymethyl cellulose columns showed an α1/α2 ratio of about 5, suggesting that at least two different species of collagen were synthesized. The α1/α2 ratio of the chains recovered from the ascorbate-deficient cultures was also about 5 but the chains were slightly underhydroxylated and the total amount of these chains which could be identified accounted for only a small fraction of the total collagen which was synthesized. A large fraction of the synthesized collagenous protein was found in the culture medium, mostly in the form of lower molecular weight peptides. It is concluded that most of the collagen which is synthesized by ascorbate-deficient tooth-bud cultures is not utilized by the component tissues, but is probably degraded and released into the medium.

Effect of ascorbic acid deficiency on mouse second molar tooth germs cultivated in vitro

Development ◽  
1976 ◽  
Vol 36 (1) ◽  
pp. 73-85
Author(s):  
Gordon E. Levenson
Keyword(s):  
Ascorbic Acid ◽  
Normal Growth ◽  
Molar Tooth ◽  
Second Molar ◽  
Root Sheath ◽  
Mandibular Second Molar ◽  
Tooth Germs ◽  
Millipore Membrane ◽  
Old Mice

Mandibular second molar tooth germs from two-day old mice were cultured in vitro, on millipore membranes, for periods of up to 20 days in liquid medium with or without added ascorbic acid. Tooth germs grown in ascorbate medium were characterized by relatively normal growth, differentiation, morphology and histology. Cuspation patterns were maintained. The epithelial root sheath continued to grow along the millipore membrane. Tooth germs cultured in ascorbate-deficient medium manifested a consistent and striking failure in maintenance of differentiated odontoblastic and ameloblastic tissue with arrest of predentin synthesis, severe structural collapse and reduction in size. Cuspation patterns were lost in scorbutic molars, with sinking of surface layers into pulpal tissue and flattening of the entire organ. This resulted in a lack of recognizable morphology and in severe disorganization of tissues. Only growing areas of the root sheath with associated proliferation of preameloblasts and pre-odontoblasts and adjacent pulpal tissue remained normal and refractory to ascorbate deficiency. Odontoblastic as well as ameloblastic layers were disrupted and cells were dedifferentiated. Newly differentiated odontoblasts became highly vacuolated when they became polarized and started to secrete extracellular matrix.

Development of embryonic rat eyes in organ culture

Development ◽  
1968 ◽  
Vol 19 (3) ◽  
pp. 407-414
Author(s):  
R. Christy Armstrong ◽  
Joel J. Elias
Keyword(s):  
Organ Culture ◽  
Culture Medium ◽  
Folic Acid Deficiency ◽  
Experimental Conditions ◽  
Acid Deficiency ◽  
Series Of Experiments ◽  
Development In Vitro ◽  
Ocular System

Abnormalities of the ocular system which appear in organ culture in Waymouth's medium with freshly added glutamine (Armstrong & Elias, 1968) resemble those caused by transitory pteryolglutamic acid (PGA or folic acid) deficiency in vivo (Armstrong & Monie, 1966). The configurations of such malformations as lens herniations, retinal diverticula, and rosette-like formations of the retina are remarkably similar in both cases. The experiments reported in this paper were undertaken in an effort to understand the mechanisms involved in the production of similar abnormalities by two very different experimental conditions: the addition of glutamine in vitro and the transitory deficiency of PGA in vivo. One series of experiments involved the effects of manipulation of the PGA and glutamine content of the culture medium on eye development in vitro. Parallel studies on PGA-deficiency in vivo were undertaken in conjunction with organ-culture experiments in order to compare the effects on abnormal eye morphogenesis.

scholarly journals Expression, localization and synthesis of small leucine-rich proteoglycans in developing mouse molar tooth germ

2020 ◽  
Vol 64 (1) ◽  
Author(s):  
Angammana Randilini ◽  
Kaoru Fujikawa ◽  
Shunichi Shibata
Keyword(s):  
Organ Culture ◽  
Developmental Stages ◽  
Tooth Development ◽  
Expression Patterns ◽  
Tooth Germ ◽  
Molar Tooth ◽  
Cervical Region ◽  
Metabolic Labeling ◽  
Tooth Formation ◽  
Tooth Germs

The gene expression and protein synthesis of small leucine-rich proteoglycans (SLRPs), including decorin, biglycan, fibromodulin, and lumican, was analyzed in the context of the hypothesis that they are closely related to tooth formation. In situ hybridization, immunohistochemistry, and organ culture with metabolic labeling of [35S] were carried out in mouse first molar tooth germs of different developmental stages using ICR mice at embryonic day (E) 13.5 to postnatal day (P) 7.0. At the bud and cap stage, decorin mRNA was expressed only in the surrounding mesenchyme, but not within the tooth germ. Biglycan mRNA was then expressed in the condensing mesenchyme and the dental papilla of the tooth germ. At the apposition stage (late bell stage), both decorin and biglycan mRNA were expressed in odontoblasts, resulting in a switch of the pattern of expression within the different stages of odontoblast differentiation. Decorin mRNA was expressed earlier in newly differentiating odontoblasts than biglycan. With odontoblast maturation and dentin formation, decorin mRNA expression was diminished and localized to the newly differentiating odontoblasts at the cervical region. Simultaneously, biglycan mRNA took over and extended its expression throughout the new and mature odontoblasts. Both mRNAs were expressed in the dental pulp underlying the respective odontoblasts. At P7.0, both mRNAs were weakly expressed but maintained their spatial expression patterns. Immunostaining showed that biglycan was localized in the dental papillae and pulp. In addition, all four SLRPs showed clear immunostaining in predentin, although the expressions of fibromodulin and lumican mRNAs were not identified in the tooth germs examined. The organ culture data obtained supported the histological findings that biglycan is more predominant than decorin at the apposition stage. These results were used to identify biglycan as the principal molecule among the SLRPs investigated. Our findings indicate that decorin and biglycan show spatial and temporal differential expressions and play their own tissue-specific roles in tooth development.

Determination of enamel protein synthesized by recombined mouse molar tooth germs in organ culture

1996 ◽  
Vol 41 (2) ◽  
pp. 215-219 ◽  
Author(s):  
Tomomi Baba ◽  
Tatsuo Terashima ◽  
Shinichiro Oida ◽  
Satoshi Sasaki
Keyword(s):  
Organ Culture ◽  
Molar Tooth ◽  
Enamel Protein ◽  
Tooth Germs

scholarly journals Rapid dissection of rodent molar-tooth germs

1981 ◽  
Vol 15 (4) ◽  
pp. 371-373 ◽  
Author(s):  
Thomas B. Kardos ◽  
Michael J. Hubbard
Keyword(s):  
Organ Culture ◽  
Direct Application ◽  
Molar Tooth ◽  
Rapid Rate ◽  
Biochemical Studies ◽  
Tooth Germs

A rapid rate of isolation of molar-tooth germs from rodents aged from 19 embryonic days to 7 days postnatal can be achieved. The procedure is of direct application to biochemical studies of odontogenesis where relatively large amounts of tissue are often required. The quality of dissection, assessed by morphological and organ culture criteria, extends the application of this procedure to general use in the isolation of molar-tooth germs from rodents.

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