concanavalin-a and cyanine-dye-3

The transport of axonal microtubules in growing neurites has been a controversial issue because of clear but conflicting results obtained with fluorescence-marking techniques. We have attempted to resolve the discordance via analysis of the relationship between apparent microtubule translocation and cell adhesion. Neuronal cultures were prepared from Xenopus embryos 1 d after injection of Cy3-conjugated tubulin into one of the blastomeres of two-cell-stage embryos. Anterograde translocation of axonal microtubules was observed in neurons cultured on a laminin-coated surface, in agreement with previously published data for Xenopus embryonic neurons. However, when neuronal cultures were prepared on a concanavalin A-treated surface, the axonal microtubules were stationary, as reported for all other neurons investigated previously. Neuronal cultures prepared on laminin- and concanavalin A-coated surfaces also demonstrated dramatic differences in the pattern of axonal growth, dynamics of axonal microtubules, and response to brefeldin A treatment. Our findings suggest that transport and dynamics of axonal microtubules may be directly affected by the mechanical tension produced by growth cone activity.

Topics: Animals; Axons; Brefeldin A; Carbocyanines; Cell Adhesion; Cell Culture Techniques; Cell Membrane; Cells, Cultured; Concanavalin A; Cyclopentanes; Fluorescent Dyes; Laminin; Microinjections; Microtubules; Neurons; Photochemistry; Protein Synthesis Inhibitors; Stress, Mechanical; Swine; Tubulin; Xenopus