Perpendicular contact guidance of CNS neuroblasts on artificial microstructures

Rodent CNS neuroblasts show parallel and perpendicular contact guidance behaviors on aligned neurite bundles in microexplant cultures (Nakatsuji, N. and Nagata, I. (1989) Development, 106, 441–447; N. I. and N. N. (1991) ibid., 112, 581–590). To test the hypothesis that the physical surface structure of the neurite bundle causes the perpendicular contact guidance, we cultured dissociated neuroblasts on quartz plates on which grating-like microstructures were fabricated by lithographic techniques. Various types of CNS neuroblasts, but not PNS neurons, oriented their processes and migrated both perpendicular and parallel to the axis of the microstructure. Perpendicular orientation was frequently observed when the microstructured grooves had depths between 0.3 micron and 0.8 micron and a width of 1 micron, which roughly mimics a tightly aligned neurite bundle. Thus, CNS neuroblasts have the ability to extend their processes and migrate perpendicular to aligned surface microstructures.

Rodent CNS neuroblasts exhibit both perpendicular and parallel contact guidance on the aligned parallel neurite bundle

Mouse cerebellar granule cells showed two types of migration behavior in microexplant cultures. They first migrated along their neurites, showing the typical contact guidance, and then oriented themselves at right angles to the parallel neurites, thus exhibiting the ‘perpendicular contact guidance’ (Nakatsuji, N. and Nagata, I. 1989 Development, 106, 441–447). To study whether other neurons have the capacity to show similar ‘perpendicular contact guidance’, we cultured dissociated neuroblasts from various parts of CNS or PNS on parallel neurite bundles. The PNS neuroblasts always extended their processes parallel to the neurite bundle. In contrast, almost all kinds of CNS neuroblasts tested oriented their processes both perpendicular and parallel to the neurite bundles that were all free of glia. Time-lapse video recording revealed that neuroblasts migrated in both directions. Thus, CNS neuroblasts possess the capacity to migrate and extend their processes at right angles to the substratum of heterotypic neurite bundles, which may play an important role in histogenesis of the CNS during development.