Assessing Attractive or Repulsive Activity of Proteins Using Hippocampal Neurons

Begin with a dish containing a silicon matrix that has multiple small channels connected to inlet and outlet ports.

Inject the test protein with repulsive activity through the inlet to fill the channels.

Incubate to allow protein adherence in the channel area, forming test protein zones, then wash.

Remove the matrix, overlay the coated area with a control protein possessing attractive activity, and incubate.

This facilitates the control protein's adherence to the empty areas between the test protein zones, creating control protein zones with an alternating striped pattern,then wash.

Add laminin to coat the striped area, then wash.

Seed the coated area with a single-cell suspension of hippocampal neurons and incubate.

In the test protein zone, the neurons interact with the proteins, triggering signals that cause the neurons to repel, inhibiting their attachment.

In contrast, neuron-protein interactions in the control protein zone promote neuronal attachment, supporting their growth and axon extension.

Prepare the fluorescently labeled recombinant proteins in PBS. Make 25 microliters for each stripe dish. Let the mixture incubate at room temperature for 30 minutes. Next, for each dis