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The Journal of Neuroscience, October 1, 1999, 19(19):8443-8453
Tenascin-C Contains Domains That Independently Regulate Neurite Outgrowth and Neurite Guidance
Sally Meiners, Mary Lynn T. Mercado, Mohammed S. A. Nur-e-Kamal, and Herbert M. Geller Department of Pharmacology, University of Medicine and Dentistry of New Jersey, Robert Wood Johnson Medical School, Piscataway, New Jersey 08854
Tenascin-C has been implicated in regulation of both neurite outgrowth and neurite guidance. We have shown previously that a particular region of tenascin-C has powerful neurite outgrowth-promoting actions in vitro. This region consists of the alternatively spliced fibronectin type-III (FN-III) repeats A-D and is abbreviated fnA-D. The purpose of this study was to investigate whether fnA-D also provides neurite guidance cues and whether the same or different sequences mediate outgrowth and guidance. We developed an assay to quantify neurite behavior at sharp substrate boundaries and found that neurites demonstrated a strong preference for fnA-D when given a choice at a poly-L-lysine-fnA-D interface, even when fnA-D was intermingled with otherwise repellant molecules. Furthermore, neurites preferred cells that overexpressed the largest but not the smallest tenascin-C splice variant when given a choice between control cells and cells transfected with tenascin-C. The permissive guidance cues of large tenascin-C expressed by cells were mapped to fnA-D. Using a combination of recombinant proteins corresponding to specific alternatively spliced FN-III domains and monoclonal antibodies against neurite outgrowth-promoting sites, we demonstrated that neurite outgrowth and guidance were facilitated by distinct sequences within fnA-D. Hence, neurite outgrowth and neurite guidance mediated by the alternatively spliced region of tenascin-C are separable events that can be independently regulated.
Key words: tenascin-C; FN-III domain; alternatively spliced region; neurite guidance; neurite outgrowth; inert substrate; cellular substrate
Copyright © 1999 Society for Neuroscience 0270-6474/99/19198443-11$05.00/0
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