Corneal collagen fibrils: Dissection with specific collagenases and monoclonal antibodies

J. M. Fitch, D. E. Birk, A. Mentzer, Karen Hasty, C. Mainardi, T. F. Linsenmayer

Research output: Contribution to journalArticle

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Abstract

To investigate the relationship of collagen types I and V within corneal fibrils, and the collagenolytic mechanisms potentially involved in corneal development and remodeling, we have incubated cryostat section of avian corneas with collagenases that specifically degrade collagen types I and V to digest selectively the collagen in situ. These preparations were then analyzed by immunofluorescence histochemistry and immunoelectron microscopy using anti-collagen type-specific monoclonal antibodies. Digestion of corneal sections with the type I-specific collagenase (''I'ase'') exposed antigenically masked type V collagen, indicating that epitopes on type V collagen in heterotypic fibrils are inaccessible to the antibody due to their interaction with type I collagen. Sections digested with the type V collagen-degrading enzyme (''V'ase'') showed no removal of type V collagen. However, after the fibrillar structure was disrupted by acetic acid treatment before enzyme digestion, the type V collagen was then degraded. Likewise, prior digestion of type I collagen by I'ase also rendered type V collagen susceptible to digestion by V'ase. These results suggest that the cleavage sites on type V collagen also are buried within heterotypic fibrils and therefore inaccessible to the enzyme. They also document, for the first time, V'ase activity against type V collagen in situ. Electron microscopic observations of sections partially digested with the I'ase revealed many short striated fibrillar segments from which smaller filaments protrude. Both the striated regions and some of the filaments were labeled by an antibody against type I collagen; anti-type V antibody reacted preferentially with the thin filaments. Thus avian corneal fibrils contain type I collagen, in which filaments of type V collagen are embedded. Complete removal of the fibrillar stromal matrix in the course of normal or pathological remodeling requires at least two different collagenases acting in concert.

Original languageEnglish (US)
Pages (from-to)1125-1136
Number of pages12
JournalInvestigative Ophthalmology and Visual Science
Volume29
Issue number7
StatePublished - Jan 1 1988

Fingerprint

Collagen Type V
Collagenases
Dissection
Collagen
Monoclonal Antibodies
Collagen Type I
Digestion
Immunoelectron Microscopy
Antibodies
Enzymes
Fluorescence Microscopy
Acetic Acid
Cornea
Epitopes
Anti-Idiotypic Antibodies

All Science Journal Classification (ASJC) codes

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

Fitch, J. M., Birk, D. E., Mentzer, A., Hasty, K., Mainardi, C., & Linsenmayer, T. F. (1988). Corneal collagen fibrils: Dissection with specific collagenases and monoclonal antibodies. Investigative Ophthalmology and Visual Science, 29(7), 1125-1136.

Corneal collagen fibrils : Dissection with specific collagenases and monoclonal antibodies. / Fitch, J. M.; Birk, D. E.; Mentzer, A.; Hasty, Karen; Mainardi, C.; Linsenmayer, T. F.

In: Investigative Ophthalmology and Visual Science, Vol. 29, No. 7, 01.01.1988, p. 1125-1136.

Research output: Contribution to journalArticle

Fitch, JM, Birk, DE, Mentzer, A, Hasty, K, Mainardi, C & Linsenmayer, TF 1988, 'Corneal collagen fibrils: Dissection with specific collagenases and monoclonal antibodies', Investigative Ophthalmology and Visual Science, vol. 29, no. 7, pp. 1125-1136.
Fitch, J. M. ; Birk, D. E. ; Mentzer, A. ; Hasty, Karen ; Mainardi, C. ; Linsenmayer, T. F. / Corneal collagen fibrils : Dissection with specific collagenases and monoclonal antibodies. In: Investigative Ophthalmology and Visual Science. 1988 ; Vol. 29, No. 7. pp. 1125-1136.
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