Binding characteristics of chondroitin sulfate proteoglycans and laminin-1, and correlative neurite outgrowth behaviors in a standard tissue culture choice assay

Diane M. Snow, Jeffrey D. Smith, Julie A. Gurwell

Research output: Contribution to journalArticlepeer-review

30 Scopus citations

Abstract

Neuronal growth cones are capable of sophisticated discrimination of environmental cues, on cell surfaces and in the extracellular matrix, to accomplish navigation during development (generation) and following nervous system injury (regeneration). Choices made by growth cones are commonly examined using tissue culture paradigms in which molecules of interest are purified and substratum-bound. From observations of growth cone behaviors using these paradigms, assertions are made about choices neuronal growth cones may make in vivo. However, in many cases, the binding, interactions, and conformations of these molecules have not been determined. In the present study, we investigated the binding characteristics of two commonly studied outgrowth regulatory molecules: Chondroitin sulfate proteoglycans (CSPGs), which are typically inhibitory to neurite outgrowth during development and following nervous system injury, and laminin, which is typically outgrowth promoting for many neuronal types. Using a novel combination of radiolabeling and quantitative fluorescence, we determined the precise concentrations of CSPGs and laminin-1 that were bound separately and together in a variety of choice assays. For identically prepared cultures, we correlated neurite outgrowth behaviors with binding characteristics. The data support our working hypothesis that neuronal growth cones are guided by the ratio of outgrowth-promoting to outgrowth-inhibiting influences in their environment, i.e., they summate local molecular cues. The response of growth cones to these molecular combinations is most likely mediated by integrins and subsequent activation of signal transduction cascades in growth cones.

Original languageEnglish
Pages (from-to)285-301
Number of pages17
JournalJournal of Neurobiology
Volume51
Issue number4
DOIs
StatePublished - Jun 15 2002

Keywords

  • Dorsal root ganglion neurons
  • H-laminin
  • Outgrowth inhibition
  • Proteoglycans
  • S-chondroitin sulfate

ASJC Scopus subject areas

  • Neuroscience (all)
  • Cellular and Molecular Neuroscience

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