Characterization of a new brain-derived proteoglycan inhibiting retinal ganglion cell axon outgrowth.
A proteoglycan was identified and isolated from physiological saline extracts of chick embryo brains by using a new monoclonal antibody (hybridoma clone mab Te38). The purified proteoglycan displayed an apparent molecular mass of 2500-3500 kDa, which became reduced to 370 and 600 kDa after digestion with chondroitinase ABC or chondroitinase AC. After additional treatment with keratanase the 600-kDa band was no longer detectable in Western blots. The specific epitope recognized by mab Te38 is an O-linked carbohydrate associated with the core protein. Tenascin-C, an extracellular matrix protein known to associate with several proteoglycans, copurified with the mab Te38 proteoglycan on the immunoaffinity column. Mab Te38 binds to the surface of nonneuronal cells; in sections from the primary visual system, expression is restricted to cells in the optic fissure, the dorsal optic nerve, and the chiasm. No retinal cells were found to express the mab Te38 epitope. The isolated molecule inhibited axon outgrowth from retinal explants when offered bound to a substrate consisting of either matrigel or collagen, chondroitinase treatment did not alter the inhibitory properties. The distribution and in vitro function of the Te38 proteoglycan indicate that it may serve a role in guidance of retinal ganglion cell axons.
Henke-Fahle
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S
Department of Ophthalmology, University of Tübingen, Germany.
Wild
K
K
Sierra
A
A
Monnier
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PP
D000818:Animals; D000911:Antibodies, Monoclonal; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D002642:Chick Embryo; D018604:Epitope Mapping; D020439:Growth Cones / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D006131:Growth Inhibitors / Q000378:metabolism*; D051379:Mice; D009900:Optic Nerve / Q000187:drug effects / Q000378:metabolism*; D011509:Proteoglycans / Q000378:metabolism*; D012160:Retina / Q000166:cytology / Q000378:metabolism*; D012165:Retinal Ganglion Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D013477:Superior Colliculi / Q000166:cytology / Q000378:metabolism*; D019063:Tenascin / Q000378:metabolism
D000911:Antibodies, Monoclonal; D006131:Growth Inhibitors; D011509:Proteoglycans; D019063:Tenascin