Chiu, Arlene Y. and Matthew, William D. and Patterson, Paul H. (1986) A monoclonal antibody that blocks the activity of a neurite regeneration-promoting factor: studies on the binding site and its localization in vivo. Journal of Cell Biology, 103 (4). pp. 1383-1398. ISSN 0021-9525. http://resolver.caltech.edu/CaltechAUTHORS:CHIjcb86
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Work from several laboratories has identified a proteoglycan complex secreted by a variety of non-neuronal cells that can promote neurite regeneration when applied to the surface of culture dishes. Using a novel immunization protocol, a monoclonal antibody (INO) was produced that blocks the activity of this outgrowth-promoting factor (Matthew, W. D., and P. H. Patterson, 1983, Cold Spring Harbor Symp. Quant. Biol. 48:625-631). We have used the antibody to analyze the components of the active site and to localize the complex in vivo. INO binding is lost when the complex is dissociated; if its components are selectively reassociated, INO binds only to a complex containing two different molecular weight species. These are likely to be laminin and heparan sulfate proteoglycan, respectively. On frozen sections of adult rat tissues, INO binding is present on the surfaces of glial cells of the peripheral, but not the central, nervous system. INO also binds to the basement membrane surrounding cardiac and skeletal muscle cells, and binding to the latter greatly increases after denervation. In the adrenal gland and kidney, INO selectively reacts with areas rich in basement membranes, staining a subset of structures that are immunoreactive for both laminin and heparan sulfate proteoglycan. In general, the outgrowth-blocking antibody binds to areas known to promote axonal regeneration and is absent from areas known to lack this ability. This suggests that this complex, which is active in culture, may be the physiological substrate supporting nerve regeneration in vivo.
|Additional Information:||Copyright © 1986 by The Rockefeller University Press. RUP grants the public the non-exclusive right to copy, distribute, or display the Work under a Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/ and http://creativecommons.org/licenses/by-nc-sa/3.0/legalcode. Received for publication 2 April 1986, and in revised form 6 June 1986. We acknowledge Josette Carnahan and Elizabeth Silvestro for their important contributions to the early histological studies, Doreen McDowell for her help with the cell culture experiments, Jami Frost for assisting with histology and photography, and William Bleisch, Karl Herrup, and Joshua Sanes for critical reading of the manuscript. The work was supported by fellowships from the American Heart Association to A.Y. Chiu and from the Jane Coffin Childs Memorial Fund to W.D. Matthew, and by grants to P.H. Patterson and W.D. Matthew from the National Institute of Neurological and Communicative Disorders and Stroke, and the McKnight Foundation.|
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