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Electron microscopy of cartilage proteoglycans.

The Histochemical Journal

|May 1, 1977

Summary

Cartilage matrix granules are proteoglycans, likely in aggregate form. Ruthenium Red staining visualizes these proteoglycans, aiding in understanding their distribution and structure in cartilage.

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Area of Science:

Biochemistry
Cell Biology
Structural Biology

Background:

Cartilage proteoglycans are complex macromolecules essential for tissue structure and function.
These proteoglycans consist of protein cores with covalently linked glycosaminoglycan chains (chondroitin sulfate and keratan sulfate).
In cartilage, proteoglycans aggregate with hyaluronic acid and link proteins to form large macromolecular complexes.

Purpose of the Study:

To identify the nature of matrix granules observed in cartilage using electron microscopy.
To investigate the distribution and structural organization of proteoglycans within the cartilaginous matrix.
To complement chemical analyses of proteoglycan structure with detailed electron microscopic observations.

Main Methods:

Electron microscopy of cartilage treated with Ruthenium Red fixative for selective matrix granule staining.
Enzymatic digestion of cartilage slices using chondroitin sulfate-degrading enzymes (hyaluronidase, chondroitinase) and a proteolytic enzyme (papain).
Chemical extraction of cartilage using 4 M guanidine-HCl.
Electron microscopy of isolated proteoglycan monomers spread on a surface with cytochrome c.

Main Results:

Ruthenium Red staining revealed dense, selective staining of matrix granules (10-50 nm) in cartilage.
Enzymatic digestion and chemical extraction significantly reduced or eliminated matrix granules and proteoglycan content.
Electron microscopy of isolated monomers showed an extended core filament with attached side-chain filaments, consistent with proteoglycan structure.

Conclusions:

Cartilage matrix granules represent proteoglycans, predominantly in aggregate form.
Ruthenium Red staining is a valuable method for studying proteoglycan distribution in thin sections.
Electron microscopy of isolated proteoglycans provides detailed structural insights that correlate with chemical data and support current models of proteoglycan organization.

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