Microlithographic determination of axonal/dendritic polarity in cultured hippocampal neurons

High resolution substrates, created using patterned self-assembled monolayers, are shown to direct axonal and dendritic process extension at the level of a single hippocampal neuron. Areas and dendrites were identified using morphological characteristics and immunocytochemical markers. Patterns were formed on glass coverslips from a co-planar monolayer of cell adhesive aminosilanes and non-adhesive fluorinated silanes. On patterned surfaces, the percentage of the total number of cells attached to the 0.71 mm(2) substrate field with compliance to the 25-mu m diameter 'somal adhesion site' reached 41 +/- 7% (mean +/- S.D., 428 cells counted). A total of 76 +/- 11% of cells that adhered to a somal attachment site developed a lone process greater than or equal to 100 mu m oriented in the direction of the continuous aminosilane pathway which was shown to express axonal markers, dells on either the fluorinated silane, which is non-permissive for neurite outgrowth, or localized on a:? aminosilane region only 5 mu m wide failed to extend major processes. This approach is amenable to a variety of industry standard fabrication techniques and may be used to study the role of fine scale spatial cues in neuronal development and synapse formation. (C) 1998 Elsevier Science B,V. All rights reserved.