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. Axons 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 mm2 substrate field with compliance to the 25-micron 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 > or = 100 microns oriented in the direction of the continuous aminosilane pathway which was shown to express axonal markers. Cells on either the fluorinated silane, which is non-permissive for neurite outgrowth, or localized on an aminosilane region only 5 microns 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.

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