Effects of cis-4-hydroxy-l-proline, an inhibitor of Schwann cell differentiation, on the secretion of collagenous and noncollagenous proteins by Schwann cells

Charles F. Eldridge, Richard P. Bunge, Mary B Bunge

Research output: Contribution to journalArticle

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Abstract

The proline analog cis-4-hydroxy-l-proline (CHP) was previously shown to inhibit both Schwann cell (SC) differentiation and extracellular matrix (ECM) formation in cultures of rat SCs and dorsal root ganglion neurons. We confirmed that CHP inhibits basal lamina formation by immunofluorescence with antibodies to laminin, type IV collagen, and heparan sulfate proteoglycan. In order to test the hypothesis that CHP inhibits SC differentiation by specifically inhibiting the secretion of collagen, cultures grown in the presence or absence of CHP were metabolically labeled with [3H]leucine and the media were analyzed for relative amounts of (a) collagenous and noncollagenous proteins by assay with bacterial collagenase and by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), or (b) triple-helical collagen by pepsin digestion followed by SDS-PAGE. The results indicate that although CHP inhibited the accumulation of secreted collagen in the culture medium and disrupted collagen triple-helix formation, it also significantly inhibited the accumulation of secreted noncollagenous proteins in the medium. CHP had no significant effect on either total protein synthesis (medium plus cell layer) or cell number. We conclude that CHP does not act as a specific inhibitor of collagen secretion in this system, and thus data from these experiments cannot be used to relate SC collagen production to other aspects of SC differentiation. We discuss the evidence for and against specificity of CHP action in other systems.

Original languageEnglish
Pages (from-to)491-501
Number of pages11
JournalExperimental Cell Research
Volume174
Issue number2
DOIs
StatePublished - Jan 1 1988
Externally publishedYes

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Schwann Cells
Proline
Cell Differentiation
Collagen
Proteins
Sodium Dodecyl Sulfate
Polyacrylamide Gel Electrophoresis
Genetic Complementation Test
Heparan Sulfate Proteoglycans
Collagen Type IV
Pepsin A
Spinal Ganglia
Laminin
Collagenases
Basement Membrane
Information Systems
Leucine
Fluorescent Antibody Technique
Extracellular Matrix
Culture Media

ASJC Scopus subject areas

  • Cell Biology

Cite this

Effects of cis-4-hydroxy-l-proline, an inhibitor of Schwann cell differentiation, on the secretion of collagenous and noncollagenous proteins by Schwann cells. / Eldridge, Charles F.; Bunge, Richard P.; Bunge, Mary B.

In: Experimental Cell Research, Vol. 174, No. 2, 01.01.1988, p. 491-501.

Research output: Contribution to journalArticle

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