Type VIII collagen has a restricted distribution in specialized extracellular matrices

R. Kapoor, Lynn Sakai, S. Funk, E. Roux, P. Bornstein, E. H. Sage

Research output: Contribution to journalArticle

58 Citations (Scopus)

Abstract

A pepsin-resistant triple helical domain (chain 50,000 M(r)) of type VIII collagen was isolated from bovine corneal Descemet's membrane and used as an immunogen for the production of mAbs. An antibody was selected for biochemical and tissue immunofluorescence studies which reacted both with Descemet's membrane and with type VIII collagen 50,000-M(r) polypeptides by competition ELISA and immunoblotting. This antibody exhibited no crossreactivity with collagen types I-VI by competition ELISA. The mAb specifically precipitated a high molecular mass component of type VIII collagen (EC2, of chain 125,000 M(r)) from the culture medium of subconfluent bovine corneal endothelial cels metabolically labeled for 24 h. In contrast, confluent cells in the presence of FCS and isotope for 7 d secreted a collagenous component of chain 60,000 M(r) that did not react with the anti-type VIII collagen IgG. Type VIII collagen therefore appears to be synthesized as a discontinuous triple helical molecule with a predominant chain 125,000 M(r) by subconfluent, proliferating cells in culture. Immunofluorescence studies with the mAb showed the type VIII collagen was deposited as fibrils in the extracellular matrix of corneal endothelial cells. In the fetal calf, type VII collagen was absent from basement membranes and was found in a limited number of tissues. In addition to the linear staining pattern observed in the Descemet's membrane, type VIII collagen was found in highly fibrillar arrays in the ocular sclera, in the meninges surrounding brain, spinal cord, and optic nerve, and in periosteum and perichondrium. Fine fibrils were evident in the white matter of spinal cord, whereas a more generalized staining was apparent in the matrices of cartilage and bone. Despite attempts to unmask the epitope, type VIII collagen was not found in aorta, kidney, lung, liver, skin, and ligament. We conclude that this unusual collagen is a component of certain specialized extracellular matrices, several of which are derived from the neural crest.

Original languageEnglish (US)
Pages (from-to)721-730
Number of pages10
JournalJournal of Cell Biology
Volume107
Issue number2
DOIs
StatePublished - Jan 1 1988
Externally publishedYes

Fingerprint

Collagen Type VIII
Extracellular Matrix
Descemet Membrane
Fluorescent Antibody Technique
Spinal Cord
Collagen Type VII
Enzyme-Linked Immunosorbent Assay
Collagen Type VI
Staining and Labeling
Meninges
Spinal Nerves
Periosteum
Sclera
Bone Matrix
Antibodies
Neural Crest
Pepsin A
Optic Nerve
Collagen Type I
Ligaments

ASJC Scopus subject areas

  • Cell Biology

Cite this

Type VIII collagen has a restricted distribution in specialized extracellular matrices. / Kapoor, R.; Sakai, Lynn; Funk, S.; Roux, E.; Bornstein, P.; Sage, E. H.

In: Journal of Cell Biology, Vol. 107, No. 2, 01.01.1988, p. 721-730.

Research output: Contribution to journalArticle

Kapoor, R. ; Sakai, Lynn ; Funk, S. ; Roux, E. ; Bornstein, P. ; Sage, E. H. / Type VIII collagen has a restricted distribution in specialized extracellular matrices. In: Journal of Cell Biology. 1988 ; Vol. 107, No. 2. pp. 721-730.
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