Type II collagen during cartilage and corneal development: immunohistochemical analysis with an anti-telopeptide antibody.
We have examined the pattern of immunoreactivity of a monoclonal antibody, II-5B2, with specificity for an epitope which resides within the NH2-terminal extension peptide (telopeptide) of the avian type II collagen molecule. This epitope is available in regions of matrix where de novo synthesis of the molecule is ongoing, but not where synthesis has ceased and maturation and crosslink formation have occurred. Within the cartilaginous growth plate, the epitope disappears from the matrix soon after the chondrocytes become hypertrophic; within the cornea, the epitope disappears subjacent to the epithelium. The II-5B2 epitope is not made available by a variety of procedures shown to remove potentially masking substances and to disrupt fibrillar organization. It is rendered available, however, when covalent crosslink formation between collagen molecules is blocked through administration of beta-aminopropionitrile or penicillamine. In contrast, the epitope of another monoclonal antibody against type II collagen, II-II6B3, which resides in the triple-helical domain of the molecule, in cartilage is present throughout the growth plate including the hypertrophic zone, and in cornea extends for a considerable distance into the stroma. Thus, it is available for antibody binding regardless of fibril maturation and crosslinking. These data suggest that the II-5B2 epitope becomes unavailable when the telopeptide becomes crosslinked. By using these two monoclonal antibodies in serial sections, one can establish the crosslinking pattern of type II collagen in the tissue.(ABSTRACT TRUNCATED AT 250 WORDS)
Chen
Q
Q
Department of Anatomy and Cellular Biology, Tufts University School of Medicine, Boston, Massachusetts 02111.
Fitch
J M
JM
Gibney
E
E
Linsenmayer
T F
TF
D000629:Aminopropionitrile; D000818:Animals; D000842:Ankle / Q000196:embryology*; D000911:Antibodies, Monoclonal; D002642:Chick Embryo; D003094:Collagen / Q000032:analysis*; D003315:Cornea / Q000196:embryology*; D000939:Epitopes / Q000032:analysis*; D005455:Fluorescent Antibody Technique; D006801:Humans; D017403:In Situ Hybridization; D010396:Penicillamine; D013977:Tibia / Q000196:embryology*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D000629:Aminopropionitrile; D003094:Collagen; D010396:Penicillamine