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N Balmain

Publications and source records attributed to N Balmain.

36 records · Page 2Linked to original sources

Calbindin-D9K immunolocalization and vitamin D-dependence in the bone of growing and adult rats.

This report presents evidence for the presence of the vitamin D-dependent calcium-binding protein, calbindin-D9K, in bone cells and matrix. In undecalcified frozen sections of growing and adult rat bone, calbindin-D9K was immunohistochemically localized in trabecular bone of the epiphysis and metaphysis and in cortical bone of the diaphysis. It was found within the cytoplasm of osteocytes, of osteoblasts lining the osteoid, and osteoblasts inside the osteoid seams. It was also found in the osteoblast processes and the anastomosed reticulum of the processes connecting the osteocytes with each other. Extracellularly, calbindin-D9K immunoreactivity was present in compact cortical bone in the areas of the mineralized matrix surrounding the osteocyte lacunae, and in the pericanalicular walls containing the cell processes. Calbindin-D9K immunoreactivity was low or absent from the cytoplasm of osteocytes in trabecular bone from severely vitamin D-deficient rats and restored in vitamin D-deficient rats given a single dose of 1,25(OH)2-VitD3. Thus, the synthesis of immunoreactive calbindin-D9K by osteoblasts and osteocytes in trabecular bone is vitamin D-dependent. The presence of immunoreactive calbindin-D9K in the osteocytes and their cell processes suggests that this calcium-binding protein is involved in the calcium fluxes regulating bone calcium homeostasis. Its localization in osteoblasts involved in bone formation and in their cell processes suggests that it has a role in the calcium transport from these cells towards the sites of active bone mineralization.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Immunological characterization, developmental pattern and vitamin-D-dependency of calbindin D-28 K in rat teeth ameloblasts.

It has been suggested that vitamin D is involved in the process of cell differentiation and extracellular mineralization during tooth development. One of the best-defined molecular markers of the action of vitamin D is a calcium-binding protein of Mr 28,000 called calbindin D-28 K (CaBP 28 K). Since this protein is present in growing teeth, we have examined its synthesis in teeth from vitamin D-replete and -deplete rats by Western blotting and immunocytochemistry with an antiserum to CaBP 28 K purified from rat kidney. The CaBP 28 K present in the enamel organ is a single molecular species migrating near 30 k Da, similarly to the kidney protein. The differentiation and maturation of odontogenic cells were followed during early postnatal development (2-12 days) in rat molars. At the light-microscope level, CaBP 28 K was only found in a single cell-type, the ameloblasts. The expression of this protein appeared to be developmentally controlled, since its distribution varied with the cell stage and the functional steps of amelogenesis. The protein was localized in the basal compartment of ameloblasts from the presecretory stage. During the early secretory stage, the concentration of cytoplasmic CaBP 28 K formed a gradient from the apical to the basal pole of the ameloblasts. Staining appeared homogeneous in the cytoplasm of later secretory ameloblasts. CaBP 28 K was discontinuously distributed during the maturation stage. This discontinuity might be related to cyclical changes in mature ameloblasts. In all stages, ameloblasts from vitamin-D-deficient rats appeared depleted of CaBP 28 K.

Ameloblasts↗

Calbindins D-9kDa and -28kDa and enamel secretion in vitamin D-deficient and control rats.

The present study focuses on the ultrastructure of enamel organ cells and the immunolocalization of calbindins D-9kDa and -28kDa during enamel secretion in Vitamin D-deficient rats. Vitamin D-deficiency disturbed the deposition of the layer of inner enamel and depleted the calbindins-content of ameloblasts. These data raise the possibility of a direct action of Vitamin D on the physiology of ameloblasts through ionic calcium homeostasis.

Ameloblasts↗

[The systemic environment and dental development. From the clinical to the molecular approach].

Clinical observation of patients with disordered phosphocalcium metabolism has demonstrated that dyschromia and/or dental dysplasias systematically accompany such disorders. A certain action of this steroid on dental buds has been demonstrated after analysis of the effects of experimental vitamin D deficiency in the rat: vitamin D would seem to control the behaviour of cells undergoing differentiation and also after this process is complete. Dentinogenesis and amelogenesis would appear principally to be affected. Two proteins, calbindins D-9K and -28K, may constitute the molecular mediators of this ameloblastic regulation.

Animals↗

Age-related changes in mineral of rat and bovine cortical bone.

The mineral of cortical bones has been studied in newborn, growing, and adult rats and in the calf and cow, using X-ray diffraction and infrared spectroscopy during the thermal decomposition of bones and by microassay of carbonate. The mineral of all the bone samples, regardless of species or age, was found to be a calcium-deficient apatite containing both CO3(2-) and HPCO4(2-) ions in the crystal lattice. The crystal size, Ca/P molar ratio, and CO3(2-) ion content of cortical bone all increased with increasing age in both the rat and the bovine. The Ca/P ratio varied from 1.51 in newborn rats to 1.69 in adults but remained that of Ca-deficient apatite even though its value was close to that of stoichiometric hydroxyapatite (1.67). Both the carbonate ion and the hydrogenophosphate ion contents varied from one animal species to another and with age within a given species. Maturation was correlated with an increase in carbonate ion content, which replaced the HPO4(2-) ions. In contrast, the calcium ion number per unit formula did not vary during maturation. Cortical bone mineral, in both species, regardless of age, can therefore be represented by the following formula: Ca8.3 (PO4)4.3 (CO3)x(HPO4)y(OH)0.3; y decreased and x increased with increasing age, (x + y) being constant, equal to 1.7.

Age Determination by Skeleton↗

Histology and microradiography of early post-natal molar tooth development in vitamin-D deficient rats.

The role of vitamin D on tooth-germ development was studied. The molars of vitamin D-deficient rats were compared with those of vitamin D-replete controls. The deficiency disturbed enamel and dentine mineralization and decreased their matrix secretion. Morphogenesis was affected; teeth were flattened and the whole of the epithelio-mesenchymal junction rippled. Where this irregularity was maximal, the inner dental epithelium and stratum intermedium were intermingled and the adjoining sub-odontoblast cells were mixed with poorly polarized odontoblasts. The cytodifferentiation of both central and sub-odontoblastic cells was inhibited. Thus vitamin D has a role in the early events of tooth development: morphogenesis, histodifferentiation and cytodifferentiation of pulp cells as well as in enamel and dentine mineralization.

Animals↗

Evidence for the presence of Calbindin-D 28K (CaBP-28K) in the tibial growth cartilages of rats.

The distribution of the vitamin-D dependent calcium-binding protein (Calbindin-D 28K) (CaBP-28K) in the tibial growth plate cartilage of the rat has been studied immunohistochemically using an antibody raised against rat renal CaBP-28K. The protein was detected mainly in the nuclei of chondrocytes and occasionally in the juxtanuclear cytoplasm. The distribution was not uniform throughout the growth plate, but concentrated in the proliferatively active chondrocytes of the resting and proliferative zones. These findings raise the possibility that CaBP-28K may be involved in the mitotic activity of the chondrocytes, acting as a regulator of the proliferative process, perhaps via intranuclear calcium.

Animals↗

Vitamin-D-dependent calcium-binding protein (CaBP-9K) in rat growth cartilage.

The presence of vitamin-D-dependent calcium-binding protein (CaBP-9K) in tibial growth-plate cartilage was immunohistochemically demonstrated using a specific antibody to rat duodenal CaBP-9K. The protein was found to be mainly localized in the cytoplasm of maturing chondrocytes. In hypertrophic chondrocytes, CaBP-9K concentrations decreased, and the protein was found in the cytoplasmic processes. No CaBP-specific immunoreactivity was seen in the hypertrophic chondrocytes of the lower calcified hypertrophic zone; in contrast, the protein was found in the extracellular lateral edges of longitudinal septa, i.e. where matrix vesicles are preferentially localized and where cartilage mineralization is initiated. These findings suggest that vitamin D has a direct function in this tissue. It also seems likely that CaBP-9K is an indicator of chondrocyte maturation, and that it is involved in the matrix vesicle-associated process of cartilage calcification.

Animals↗

[Molecular approach to the action of vitamin D in man].

Some applications to man of specific markers of the molecular action of vitamin D (1.25(OH)2D3 receptors and antibodies to hormone-dependent proteins (CaBP and cDNA] are reported in this study. On case of type II vitamin-dependent rickets was characterized by 1.25(OH)2D3 plasma level greater than 250 pg/ml and a ten-fold decrease of the number of binding sites of the hormone in cultured skin fibroblasts. We propose that CaBP 28K and/or 9K-containing cells, such as Purkinje's cells and chondroblasts may be targets for vitamin D action. Detection in fetuses, from the 20th week of gestation, of CaBP 9K messenger RNA in the duodenum and sternum and presence of CaBP 28K and 9K in the chondroblasts of the upper extremity of tibia, suggest that vitamin D acts on the nucleus of its target-cells during fetal development. Finally, discovery of the gene of CaBP 9K in man opens the prospect of studies which will improve the understanding of the mechanism of action of vitamin D.

Adolescent↗

Influence of starvation on sucrase regulation by dietary sucrose in the rat.

We have studied the action of sucrose on jejunal sucrase activity. Rats (175 g) were first starved or fed a digestible carbohydrate-free diet for 60 h and then fed a high sucrose diet for varying times up to 84 h. 1) Rats starved for 60 h showed mucosal atrophy with a decrease in protein content/10 cm (18.00 +/- 1.4 versus 40.1 +/- 3 mg (controls p less than 0.001) and in villus height (357 +/- 18 versus 526 +/- 5 microns, p less than 0.001) which was fully repaired only after 60 h on the sucrose diet (528 +/- 11 microns). Rats on digestible carbohydrate-free diet showed no mucosal atrophy. 2) Starved rats had a delayed (60 h) sucrase activity response to sucrose (53 +/- 7 versus 122 +/- 4 microns/mg protein, p less than 0.001). Maximum activity was obtained after 12 h on sucrose diet in rats maintained on the carbohydrate-free diet: 38 +/- 1 versus 108 +/- 2.3 microns/mg protein, p less than 0.001. 3) Villus and crypt cell analysis after starvation and 12 h on a high sucrose diet localized the increase in sucrase activity to the villus-crypt junction. No change occurred in the upper villus. The increase was complete all along the villus by 36 h. In contrast, after the carbohydrate-free diet, sucrase activity increased maximally at all levels of the villus by 12 h on the high sucrose diet.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Lung mechanics in rachitic rats.

Lung mechanics was studied at 50 days of age in 7 rachitic rats born from mothers deprived of vitamin D. They were compared with 7 control rats raised in the same conditions but fed a diet supplemented with vitamin D. The animals were anesthetized, tracheotomized, and paralyzed. Quasi-static pressure-volume curves of the respiratory system and of the lungs were obtained. Body weight of the rachitic rats was within the range of the control rats, but dry lung weight (LW) was significantly lower (p less than 0.01). Lung volumes in absolute terms and when normalized for LW were significantly lower than in the control rats. Chest wall compliance (Ccw) of the rachitic rats was within the range of values of the control rats, except for 2 animals with an infinite Ccw. Analysis of the pressure-volume curves of the lungs of the rachitic rats compared with those in the control animals showed a significant decrease in lung compliance (CL) and in CL/LW (p less than 0.01), indicating a decrease in lung distensibility. The more severe the rickets (according to microradiographic criteria of the tibia), the lower the CL/LW. It is speculated that decrease in lung distensibility may be related to abnormal lung growth caused by disturbed alveolar formation and lung connective tissue development. These abnormalities could be due to vitamin D deficiency acting on the growing lung, as on the growing bones, by a mechanism involving proteoglycans.

Animals↗

Metaphyseal pattern: uniqueness of this structure in growing bones originating from cartilaginous anlage. A microradiographic study.

Microradiographic examination of metaphyses in long and short tubular growing bones allowed detection of a repetitive, clearly defined pattern of three adjacent zones; the latter are successively formed by the mineralization of cartilaginous longitudinal intercolumnar septa and by the subsequent apposition of other mineralized tissues concurrently with resorption. Consequently, each zone of the metaphysis includes mineralized tissues of various compositions and ages, identifiable by their different mineralization densities. Microradiography of pieces of the growing skeleton in several animal species shows that the same organization is not only present in long and short tubular bones but also in many other such as the pelvis and scapula, cuboid bones like the calcaneum and talus, and cartilaginous bones at the base of the skull. This suggests that there is no difference between the osteogenesis pattern of these bones and tubular ones. The problem of identifying the factors generating such metaphyseal organization is raised.

Animals↗

The mineralized ring, a single structure peculiar to long bone growth.

Mineralized ring, an elongated tubular structure ensheathing bone metaphysis, is described at successive stages of long bone growth. It is shown that the mineralized ring is characterized by different morphological aspects during its development, corresponding to successive growth stages of one and the same anatomical formation. Its origin is different from periosteal bone and its development is parallel to that of the metaphysis.

Animals↗

X-ray diffraction of calcined bone tissue: a reliable method for the determination of bone Ca/P molar ratio.

A method is proposed which allows direct measurement by X-ray diffraction of the Ca/P molar ratio in bone tissue after calcination at 900 degrees C. It is based on the construction of a standard curve from known mixtures of hydroxyapatite (HA) and beta-tricalcium phosphate (beta-TP), since these are the only two crystallized mineral phases in the bone ash after its calcination at 900 degrees C. This method, based on physical analysis, may also be used to calculate the respective proportions of calcium and phosphorus after the Ca/P molar ratio has been obtained. It is more sensitive than and at least as accurate as the usual chemical techniques, even though it can be applied to minute samples of bone tissue. Furthermore, the method permits simultaneous determination of lattice parameters and provides additional information about bone mineral structure.

Animals↗