Effect of trifluoperazine and other drugs on matrix vesicle formation by chicken growth plate chondrocytes in primary cell culture

Schalk, Elaine M.; Wuthier, Roy E.

doi:10.1016/0006-2952(86)90464-8

Cited by 7 publications

(4 citation statements)

References 25 publications

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“…This reveals that MVs form by mechanisms that involve depolymerization of actin filaments. On the other hand, studies using tricyclic amine drugs that selectively bind to the annexins also stimulate release of MeVs from cultured chondrocytes (70). Taken together, these findings indicate that processes that uncouple the annexins from the cytoskeletal network must be involved in MV formation.…”

Section: Cell Culture Studiesmentioning

confidence: 86%

“…The use of growth plate chondrocyte cultures that release vesicles into the culture medium was also used to gain some insight into the protein and lipid composition of cell-released vesicles and in the mechanism of MV formation (68)(69)(70)(71)(72). However, a significant problem was that vesicles released into the culture medium (MeV) are not identical to those trapped in the matrix (71,(73)(74)(75).…”

Section: Cell Culture Methodsmentioning

confidence: 99%

“…Several early studies explored the sodium dodecyl sulfate -polyacrylamide gel electrophoresis (SDS-PAGE) patterns of proteins present in MV using crude collagenase digestion (88)(89)(90), as well as MVEM (55-57, 65, 68) and culture media vesicles (58,69,70,91). While there was considerable variability in the patterns of MV proteins from the various tissues, animal species, and MV preparations, certain protein bands nevertheless were consistently observed; these include two or three intensely staining bands in the 30-38 kDa range, two strongly staining bands at 40-48 kDa, and bands frequently appearing in the 55 kDa, 90-100 kDa and 130-150 kDa range.…”

Section: Early Sds-page Studiesmentioning

confidence: 99%

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Matrix vesicles: structure, composition, formation and function in calcification

Wuthier

Lipscomb²

2011

Front Biosci

158

230

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Matrix vesicles (MVs) induce calcification during endochondral bone formation. Experimental methods for structural, compositional, and functional analysis of MVs are reviewed. MV proteins, enzymes, receptors, transporters, regulators, lipids and electrolytes are detailed. MV formation is considered from both structural and biochemical perspectives. Confocal imaging of Ca(2+) and H(+) were used to depict how living chondrocytes form MVs. Biochemical studies revealed that coordinated mitochondrial Ca(2+) and Pi metabolism produce MVs containing a nucleational complex (NC) of amorphous calcium phosphate, phosphatidylserine and annexin A5--all critical to the mechanism of mineral nucleation. Reconstitution of the NC and modeling with unilamellar vesicles reveal how the NC transforms into octacalcium phosphate, regulated by Mg(2+), Zn(2+) and annexin A5. Extravasation of intravesicular mineral is mediated by phospholipases and tissue-nonspecific alkaline phosphatase (TNAP). In the extravesicular matrix, hydroxyapatite crystal propagation is enhanced by cartilage collagens and TNAP, which destroys inhibitory PPi, and by metalloproteases that degrade proteoglycans. Other proteins also modulate mineral formation. Recent findings from single and multiple gene knockouts of TNAP, NPP1, ANK, PHOSPHO1, and Annexin A5 are reviewed.

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Section: Cell Culture Studiesmentioning

confidence: 86%

Section: Cell Culture Methodsmentioning

confidence: 99%

Section: Early Sds-page Studiesmentioning

confidence: 99%

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Matrix vesicles: structure, composition, formation and function in calcification

Wuthier

Lipscomb²

2011

Front Biosci

158

230

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“…As an alternative to growth plate cartilage, chondrocyte cultures [74,86,[109][110][111][112][113][114][115][116][117][118][119][120][121][122][123] can be used as a starting material to compare the properties of media vesicles and matrix vesicles. Chondrocytes isolated from the proliferating and hypertrophic regions of the growth plate differ significantly in their ability to release TNAP-rich matrix vesicles [105].…”

Section: Matrix Vesicles and Media Vesicles From Growth Plate Cartila...mentioning

confidence: 99%

Do Media Extracellular Vesicles and Extracellular Vesicles Bound to the Extracellular Matrix Represent Distinct Types of Vesicles?

Mebarek,

Buchet,

Pikula

et al. 2023

Biomolecules

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Mineralization-competent cells, including hypertrophic chondrocytes, mature osteoblasts, and osteogenic-differentiated smooth muscle cells secrete media extracellular vesicles (media vesicles) and extracellular vesicles bound to the extracellular matrix (matrix vesicles). Media vesicles are purified directly from the extracellular medium. On the other hand, matrix vesicles are purified after discarding the extracellular medium and subjecting the cells embedded in the extracellular matrix or bone or cartilage tissues to an enzymatic treatment. Several pieces of experimental evidence indicated that matrix vesicles and media vesicles isolated from the same types of mineralizing cells have distinct lipid and protein composition as well as functions. These findings support the view that matrix vesicles and media vesicles released by mineralizing cells have different functions in mineralized tissues due to their location, which is anchored to the extracellular matrix versus free-floating.

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Effect of amino acid levels on matrix vesicle formation by epiphyseal growth plate chondrocytes in primary culture

Ishikawa

Chin

Schalk

et al. 1986

Journal Cellular Physiology

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The effect of varying the amino acid concentrations of the culture medium on matrix vesicle formation was studied in primary cultures of chicken epiphyseal growth plate chondrocytes grown in Dulbecco's modified Eagle's medium (DME) supplemented with 10% fetal bovine serum (FBS). Decreasing the levels of free amino acids in the culture medium to levels of one-half, one quarter, and one eighth of the values normally present in DME caused a progressive decline in matrix vesicle (MV) formation. Increasing the level in the culture medium of those amino acids that are enriched in extracellular fluid (ECF) of growth plate cartilage significantly increased formation of matrix vesicles (MV), as assayed by the alkaline phosphatase (AP) activities present in high-speed sediments from spent culture media. However, adjusting the levels of all amino acids to match those of the ECF produced the greatest stimulation of MV formation. Of the amino acids that are notably enriched in ECF, glutamate (GLU), alanine (ALA), serine (SER), asparagine (ASN), and taurine (TAU) individually enhanced MV production, whereas proline (PRO), glycine (GLY), and aspartate (ASP) had essentially no effect. The simple combination of ECF levels of ALA and GLU resulted in a stimulation of MV formation equal to that observed when the eight aforementioned amino acids were elevated to ECF levels. Other combinations of ASP and GLY, or of TAU, SER, and ASN showed some stimulation, but at a lower level. Increasing the amino acid concentrations, alone or in combination, also increased the levels of cellular AP, and to a lesser extent cellular protein. While increases in cellular AP were generally correlated with increased formation of AP-rich MV, this was not uniformly true. These results indicate that in addition to hormones and growth factors, nutritional factors such as the levels of amino acids are also critical for normal phenotypic expression, growth, and matrix formation by epiphyseal chondrocytes.

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Effect of trifluoperazine and other drugs on matrix vesicle formation by chicken growth plate chondrocytes in primary cell culture

Cited by 7 publications

References 25 publications

Matrix vesicles: structure, composition, formation and function in calcification

Matrix vesicles: structure, composition, formation and function in calcification

Do Media Extracellular Vesicles and Extracellular Vesicles Bound to the Extracellular Matrix Represent Distinct Types of Vesicles?

Effect of amino acid levels on matrix vesicle formation by epiphyseal growth plate chondrocytes in primary culture

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