THE EFFECT OF A SLIGHTLY ACIDIC SOMATOMEDIN PEPTIDE (ILAs) ON THE SULPHATION OF PROTEOGLYCANS FROM ARTICULAR AND GROWTH PLATE CHONDROCYTES IN CULTURE

1978 ◽  
Vol 89 (2) ◽  
pp. 263-275 ◽  
Author(s):  
Marie-Thérèse Corvol ◽  
Marie-France Dumontier ◽  
Raphael Rappaport ◽  
Harvey Guyda ◽  
Barry I. Posner
Keyword(s):  
Growth Plate ◽  
Deae Cellulose ◽  
Morphological Characteristics ◽  
Cell Types ◽  
Epiphyseal Growth Plate ◽  
Growth Plate Chondrocytes ◽  
Electrophoretic Patterns ◽  
Agarose Electrophoresis ◽  
Well Differentiated

ABSTRACT Chondrocyte cultures were prepared from rabbit growth plate (GPC) and articular (ARC) chondrocytes. These two cell types have distinct morphological characteristics. The cells reached maximum numbers by days 10 and 21 for ARC and GPC, respectively. The proteoglycans (PG) contained in the cellular pool were extracted and purified by DEAE cellulose chromatography. The effect of a partially purified somatomedin peptide with insulin-like activity on [35S] sulphate incorporation into PG was evaluated. In both ARC and GPC a significant stimulation of [35S]sulphate uptake into PG subunits was obtained with 1 ng Eq./ml of somatomedin peptide. In order to obtain the same stimulatory effect with porcine insulin, a 1000-fold greater concentration was required. The electrophoretic patterns of the PG subunits on acrylamide-agarose electrophoresis were identical on control incubations and after stimulation with the somatomedin peptide. These data demonstrate in vitro biological activity of this peptide on well differentiated articular and epiphyseal growth plate chondrocytes in culture. These cultures appear to provide a sensitive biological assay for somatomedin peptides.

TRPM7 channels mediate spontaneous Ca2+ fluctuations in growth plate chondrocytes that promote bone development

2019 ◽  
Vol 12 (576) ◽  
pp. eaaw4847 ◽  
Author(s):  
Nianchao Qian ◽  
Atsuhiko Ichimura ◽  
Daisuke Takei ◽  
Reiko Sakaguchi ◽  
Akihiro Kitani ◽  
...  
Keyword(s):  
Growth Plate ◽  
Transient Receptor Potential ◽  
Ex Vivo ◽  
Bone Development ◽  
Receptor Potential ◽  
Epiphyseal Growth Plate ◽  
Growth Plate Chondrocytes ◽  
Transient Receptor ◽  
Ex Vivo Culture

During endochondral ossification of long bones, the proliferation and differentiation of chondrocytes cause them to be arranged into layered structures constituting the epiphyseal growth plate, where they secrete the cartilage matrix that is subsequently converted into trabecular bone. Ca2+ signaling has been implicated in chondrogenesis in vitro. Through fluorometric imaging of bone slices from embryonic mice, we demonstrated that live growth plate chondrocytes generated small, cell-autonomous Ca2+ fluctuations that were associated with weak and intermittent Ca2+ influx. Several genes encoding Ca2+-permeable channels were expressed in growth plate chondrocytes, but only pharmacological inhibitors of transient receptor potential cation channel subfamily M member 7 (TRPM7) reduced the spontaneous Ca2+ fluctuations. The TRPM7-mediated Ca2+ influx was likely activated downstream of basal phospholipase C activity and was potentiated upon cell hyperpolarization induced by big-conductance Ca2+-dependent K+ channels. Bones from embryos in which Trpm7 was conditionally knocked out during ex vivo culture exhibited reduced outgrowth and displayed histological abnormalities accompanied by insufficient autophosphorylation of Ca2+/calmodulin-dependent protein kinase II (CaMKII) in the growth plate. The link between TRPM7-mediated Ca2+ fluctuations and CaMKII-dependent chondrogenesis was further supported by experiments with chondrocyte-specific Trpm7 knockout mice. Thus, growth plate chondrocytes generate spontaneous, TRPM7-mediated Ca2+ fluctuations that promote self-maturation and bone development.

scholarly journals In vivo and in vitro Chondrocyte Metabolism in Relationship to the Developemnt of Tibial Dyschondroplasia in Broiler Chickens

1993 ◽  
Author(s):  
Roland M. Leach ◽  
Mark Pines ◽  
Carol V. Gay ◽  
Shmuel Hurwitz
Keyword(s):  
In Situ Hybridization ◽  
Growth Plate ◽  
Bone Growth ◽  
Future Research ◽  
Epiphyseal Growth Plate ◽  
Growth Plate Chondrocytes ◽  
Tibial Dyschondroplasia ◽  
Sequence Of Events

Skeletal deformities are a significant financial and welfare problem for the world poultry industry. Tibial dyschondroplasia (TD) is the most prevalent skeletal abnormality found in young broilers, turkeys and ducks. Tibial dyschondroplasia results from a perturbation of the sequence of events in the epiphyseal growth plate, the tissue responsible for longitudinal bone growth. The purpose of this investigation was to test the hypothesis that TD was the result of a failure of growth plate chondrocytes to differentiate and express the chemotactic molecules required for cartilage vascularization. In this investigation in situ hybridization and immunocytochemical techniques were used to study chondrocyte gene products associated with cartilage maturation and vascularization such as osteopontin, osteonectin, type X collagen, and alkaline phosphatase. All markers were present in the growth plate tissue anter or to the TD lesion but were greatly diminished in the TD lesion. Thus, rather than not acquiring the markers for hypertrophy, it appears that the growth plate chondrocytes reach a certain stage of hypertrophy and then de-differentiate into cells which resemble chondrocytes in the prehypertrophic zone. Similar patterns were observed in all TD tissues examined whether the lesions were spontaneous or induced by dietary treatments or genetic selection. The decrease in gene expression can at least be partially explained by the fact that many of the dysplastic chondrocytes show classic signs of apoptosis. These results provide an explanation for the observation that a variety of genes show reduced expression in the TD lesion when examined by in situ hybridization. This would suggest that future research should focus on the earliest detectable stages in the development of TD and examine endocrine and autocrine factors which cause chondrocytes to de-differentiate and undergo premature apoptosis.

scholarly journals Peroxisome Proliferator-Activated Receptor-γPromotes Adipogenic Changes in Growth Plate Chondrocytes In Vitro

PPAR Research ◽  
2006 ◽  
Vol 2006 ◽  
pp. 1-8 ◽  
Author(s):  
Lai Wang ◽  
Yvonne Y. Shao ◽  
R. Tracy Ballock
Keyword(s):  
Growth Plate ◽  
Adipogenic Differentiation ◽  
Terminal Differentiation ◽  
Cell Types ◽  
Peroxisome Proliferator ◽  
Growth Plate Chondrocytes ◽  
Peroxisome Proliferator Activated Receptor ◽  
Infected Cells ◽  
In Vitro Data

Chondrocytes and adipocytes are two differentiated cell types which are both derived from mesenchymal cells. The purpose of this study was to investigate whether peroxisome proliferator-activated receptor-γ(PPARγ), a transcription factor involved in lineage determination during adipogenesis, is able to induce adipogenic differentiation in growth plate chondrocytes. Isolated epiphyseal chondrocytes were infected with a PPARγadenovirus or treated with the PPARγagonist ciglitazone. Both of these treatments resulted in lipid droplet accumulation and expression of the adipogenic markers aP2, lipoprotein lipase, and adipsin in chondrocytes. Proteoglycan matrix synthesis was decreased in the PPARγ-infected cells, as was the expression of the chondrogenic genes Col2a1 and aggrecan. Growth plate cells transfected with a PPARγexpression plasmid under the control of the collagenα1(II) promoter also demonstrated a similar adipogenic changes. Terminal differentiation of growth plate chondrocytes induced by thyroid hormone was also inhibited by overexpression of PPARγand ciglitazone treatment, with decreased expression of alkaline phosphatase and Runx2/Cbfa1 genes. These in vitro data suggest that PPARγis able to promote adipogenic differentiation in growth plate chondrocytes, while negatively regulating chondrogenic differentiation and terminal differentiation.

HiPER1, a phosphatase of the endoplasmic reticulum with a role in chondrocyte maturation

1998 ◽  
Vol 111 (6) ◽  
pp. 803-813
Author(s):  
P.R. Romano ◽  
J. Wang ◽  
R.J. O'Keefe ◽  
J.E. Puzas ◽  
R.N. Rosier ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Growth Plate ◽  
Articular Chondrocytes ◽  
Signal Sequence ◽  
Growth Plate Chondrocytes ◽  
Chondrocyte Maturation ◽  
Er Retention ◽  
Alternatively Spliced ◽  
Dual Label

We have previously identified and partially cloned Band 17, a gene expressed in growth plate chondrocytes transiting from proliferation to hypertrophy. We now rename this gene HiPER1, Histidine Phosphatase of the Endoplasmic Reticulum-1, based on the results reported here. HiPER1 encodes two proteins of 318 (HiPER1(318)) and 449 (HiPER1(449)) amino acids, which are 20–21% identical to a group of yeast acid phosphatases that are in the histidine phosphatase family. HiPER1(449) is significantly more abundant than HiPER1(318), correlating with the abundance of the alternatively spliced messages encoding HiPER449 and HiPER318. Anti-HiPER1 antibodies detect two proteins of 53 and 55 kDa in growth plate chondrocytes that are absent in articular chondrocytes. We confirm that the 53 and 55 kDa proteins are HiPER1(449) by heterologous expression of the HiPER1(449) coding sequence in chick embryo fibroblasts. The 53 and 55 kDa proteins are glycosylated forms of HiPER1(449), as N-glycosidase F digestion reduces these proteins to 48 kDa, the predicted size of HiPER1(449) without the N-terminal signal sequence. Immunocytochemistry demonstrates that HiPER1(449) is found in chondrocytes maturing from proliferation to hypertrophy, but is not detectable in resting zone, deep hypertrophic zone or articular chondrocytes, a distribution that is consistent with the message distribution. HiPER1(449) was predicted to localize to the lumen of endoplasmic reticulum by an N-terminal signal sequence and by the C-terminal sequence Ala-Asp-Glu-Leu, which closely matches the consensus signal for ER retention, Lys-Asp-Glu-Leu. We confirm this prediction by demonstrating colocalization of HiPER1(449) with the ER protein HSP47 using dual-label immunofluorescence. PTHrP, a peptide that prevents hypertrophy in chondrocytes, suppressed HiPER1 and HiPER1(449) expression in vitro, an observation that further supports a role for HiPER1 in chondrocyte maturation. The yeast phosphatase homology, localization to the endoplasmic reticulum and pattern of expression suggest that HiPER1 represents a previously unrecognized intracellular pathway, involved in differentiation of chondrocytes.

scholarly journals Parathyroid hormone prevents 1,25(OH)2D3 induced down-regulation of the vitamin D receptor in growth plate chondrocytes in vitro

1997 ◽  
Vol 52 (1) ◽  
pp. 45-51 ◽  
Author(s):  
Günter Klaus ◽  
Tanja May ◽  
Ulrike Hügel ◽  
Barbara Von Eichel ◽  
Julian Rodriguez ◽  
...  
Keyword(s):  
Vitamin D ◽  
Parathyroid Hormone ◽  
Growth Plate ◽  
Vitamin D Receptor ◽  
Growth Plate Chondrocytes ◽  
Down Regulation

scholarly journals Type B γ-Aminobutyric Acid Receptors Modulate the Function of the Extracellular Ca2+-Sensing Receptor and Cell Differentiation in Murine Growth Plate Chondrocytes

Endocrinology ◽  
2007 ◽  
Vol 148 (10) ◽  
pp. 4984-4992 ◽  
Author(s):  
Zhiqiang Cheng ◽  
Chialing Tu ◽  
Luis Rodriguez ◽  
Tsui-Hua Chen ◽  
Melita M. Dvorak ◽  
...  
Keyword(s):  
Cell Differentiation ◽  
Growth Plate ◽  
Type Ii Collagen ◽  
Aminobutyric Acid ◽  
Physical Interaction ◽  
Type B ◽  
Growth Plate Chondrocytes ◽  
Differentiation Markers ◽  
Receptor Complexes

Extracellular calcium-sensing receptors (CaRs) and metabotropic or type B γ-aminobutyric acid receptors (GABA-B-Rs), two closely related members of family C of the G protein-coupled receptor superfamily, dimerize in the formation of signaling and membrane-anchored receptor complexes. We tested whether CaRs and two GABA-B-R subunits (R1 and R2) are expressed in mouse growth plate chondrocytes (GPCs) by PCR and immunocytochemistry and whether interactions between these receptors influence the expression and function of the CaR and extracellular Ca2+-mediated cell differentiation. Both CaRs and the GABA-B-R1 and -R2 were expressed in the same zones of the growth plate and extensively colocalized in intracellular compartments and on the membranes of cultured GPCs. The GABA-B-R1 coimmunoprecipitated with the CaR, confirming a physical interaction between the two receptors in GPCs. In vitro knockout of GABA-B-R1 genes, using a Cre-lox recombination strategy, blunted the ability of high extracellular Ca2+ concentration to activate phospholipase C and ERK1/2, suppressed cell proliferation, and enhanced apoptosis in cultured GPCs. In GPCs, in which the GABA-B-R1 was acutely knocked down, there was reduced expression of early chondrocyte markers, aggrecan and type II collagen, and increased expression of the late differentiation markers, type X collagen and osteopontin. These results support the idea that physical interactions between CaRs and GABA-B-R1s modulate the growth and differentiation of GPCs, potentially by altering the function of CaRs.

scholarly journals Expression of tissue transglutaminase in skeletal tissues correlates with events of terminal differentiation of chondrocytes.

1993 ◽  
Vol 120 (6) ◽  
pp. 1461-1470 ◽  
Author(s):  
D Aeschlimann ◽  
A Wetterwald ◽  
H Fleisch ◽  
M Paulsson
Keyword(s):  
Growth Plate ◽  
Tissue Transglutaminase ◽  
Specific Substrate ◽  
Epiphyseal Growth Plate ◽  
Calcified Cartilage ◽  
Tissue Homogenates ◽  
The One ◽  
Bone Trabeculae ◽  
Rat Paw

Calcifying cartilages show a restricted expression of tissue transglutaminase. Immunostaining of newborn rat paw bones reveals expression only in the epiphyseal growth plate. Tissue transglutaminase appears first intracellularly in the proliferation/maturation zone and remains until calcification of the tissue in the lower hypertrophic zone. Externalization occurs before mineralization. Subsequently, the enzyme is present in the interterritorial matrix during provisional calcification and in the calcified cartilage cores of bone trabeculae. In trachea, mineralization occurring with maturation in the center of the cartilage is accompanied by expression of tissue transglutaminase at the border of the hydroxyapatite deposits. Transglutaminase activity also shows a restricted distribution in cartilage, similar to the one observed for tissue transglutaminase protein. Analysis of tissue homogenates showed that the enzyme is present in growth plate cartilage, but not in articular cartilage, and recognizes a limited set of substrate proteins. Osteonectin is coexpressed with tissue transglutaminase both in the growth plate and in calcifying tracheal cartilage and is a specific substrate for tissue transglutaminase in vitro. Tissue transglutaminase expression in skeletal tissues is strictly regulated, correlates with chondrocyte differentiation, precedes cartilage calcification, and could lead to cross-linking of the mineralizing matrix.

Sign in / Sign up

Export Citation Format

Share Document