Da. Stenger et al., MICROLITHOGRAPHIC DETERMINATION OF AXONAL DENDRITIC POLARITY IN CULTURED HIPPOCAMPAL-NEURONS/, Journal of neuroscience methods, 82(2), 1998, pp. 167-173
High resolution substrates, created using patterned self-assembled mon
olayers, are shown to direct axonal and dendritic process extension at
the level of a single hippocampal neuron. Areas and dendrites were id
entified using morphological characteristics and immunocytochemical ma
rkers. Patterns were formed on glass coverslips from a co-planar monol
ayer of cell adhesive aminosilanes and non-adhesive fluorinated silane
s. 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-m
u m diameter 'somal adhesion site' reached 41 +/- 7% (mean +/- S.D., 4
28 cells counted). A total of 76 +/- 11% of cells that adhered to a so
mal attachment site developed a lone process greater than or equal to
100 mu m oriented in the direction of the continuous aminosilane pathw
ay which was shown to express axonal markers, dells on either the fluo
rinated silane, which is non-permissive for neurite outgrowth, or loca
lized on a:? aminosilane region only 5 mu m wide failed to extend majo
r processes. This approach is amenable to a variety of industry standa
rd fabrication techniques and may be used to study the role of fine sc
ale spatial cues in neuronal development and synapse formation. (C) 19
98 Elsevier Science B,V. All rights reserved.