Characterization of an in vitro model of elastic fiber assembly

Bruce Robb, Hiroshi Wachi, Theresa Schaub, Robert P. Mecham, Elaine C. Davis

Research output: Contribution to journalArticle

55 Citations (Scopus)

Abstract

Elastic fibers consist of two morphologically distinct components: elastin and 10-nm fibrillin-containing micro fibrils. During development, the micro fibrils form bundles that appear to act as a scaffold for the deposition, orientation, and assembly of tropoelastin monomers into an insoluble elastic fiber. Although microfibrils can assemble independent of elastin, tropoelastin monomers do not assemble without the presence of microfibrils. In the present study, immortalized ciliary body pigmented epithelial (PE) cells were investigated for their potential to serve as a cell culture model for elastic fiber assembly. Northern analysis showed that the PE cells express microfibril proteins but do not express tropoelastin. Immunofluorescence staining and electron microscopy confirmed that the microfibril proteins produced by the PE cells assemble into intact microfibrils. When the PE cells were transfected with a mammalian expression vector containing a bovine tropoelastin cDNA, the cells were found to express and secrete tropoelastin. Immunofluorescence and electron microscopic examination of the transfected PE cells showed the presence of elastic fibers in the matrix. Biochemical analysis of this matrix showed the presence of cross-links that are unique to mature insoluble elastin. Together, these results indicate that the PE cells provide a unique, stable in vitro system in which to study elastic fiber assembly.

Original languageEnglish (US)
Pages (from-to)3595-3605
Number of pages11
JournalMolecular Biology of the Cell
Volume10
Issue number11
StatePublished - Nov 1999
Externally publishedYes

Fingerprint

Tropoelastin
Elastic Tissue
Microfibrils
Epithelial Cells
Elastin
Ciliary Body
Fluorescence Microscopy
Fluorescent Antibody Technique
In Vitro Techniques
Electron Microscopy
Proteins
Complementary DNA
Cell Culture Techniques
Electrons
Staining and Labeling

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Robb, B., Wachi, H., Schaub, T., Mecham, R. P., & Davis, E. C. (1999). Characterization of an in vitro model of elastic fiber assembly. Molecular Biology of the Cell, 10(11), 3595-3605.

Characterization of an in vitro model of elastic fiber assembly. / Robb, Bruce; Wachi, Hiroshi; Schaub, Theresa; Mecham, Robert P.; Davis, Elaine C.

In: Molecular Biology of the Cell, Vol. 10, No. 11, 11.1999, p. 3595-3605.

Research output: Contribution to journalArticle

Robb, B, Wachi, H, Schaub, T, Mecham, RP & Davis, EC 1999, 'Characterization of an in vitro model of elastic fiber assembly', Molecular Biology of the Cell, vol. 10, no. 11, pp. 3595-3605.
Robb B, Wachi H, Schaub T, Mecham RP, Davis EC. Characterization of an in vitro model of elastic fiber assembly. Molecular Biology of the Cell. 1999 Nov;10(11):3595-3605.
Robb, Bruce ; Wachi, Hiroshi ; Schaub, Theresa ; Mecham, Robert P. ; Davis, Elaine C. / Characterization of an in vitro model of elastic fiber assembly. In: Molecular Biology of the Cell. 1999 ; Vol. 10, No. 11. pp. 3595-3605.
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