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. 2015 Jul 14;9:32. doi: 10.3389/fncir.2015.00032

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Copyright © 2015 Pan, Alagapan, Franca, Leondopulos, DeMarse, Brewer and Wheeler.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution and reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Timed sequential plating of cortical neurons for feed-forward connectivity. Each micro-tunnel device was aligned and attached to a 60 electrode MEA. Cortical neurons were first seeded in Well A on Day 0. During the following 6 days neurites extend from soma in Well A and synapse with other soma in Well A while others enter the tunnels and extend into the adjacent well. Eventually enough axons have entered the tunnels that on Day 7 when additional neurons are added to Well B there is little room for neurites from those new neurons to cross back into Well A. The result is a primarily feed-forward network in which the majority of cross-well connectivity is biased in one direction (Well A to Well B). In this paper we manipulate the number of tunnels that connect each chamber and measure the effect this has on the “strength” of the connection between two neural populations separated by these tunnels.