The cortical circuitry in the brain consists of structurally and functionally distinct neuronal assemblies with reciprocal axon connections. perform mechanistic studies. Micro-patterned surfaces have been used in neuronal cultures to fabricate neural networks with pre-determined geometry. The patterns provide selective surfaces to control neuronal attachment and growth (Jung et al., 2001, Li et al., 2003,Corey and Feldman., 2003). Surface patterning methods include microfluidic channels (Taylor et al., 2005), laser-ablated microgrooves (Corey et al., 1991), silane coupling-mediated covalent binding (Kleinfeld et al., 1988,Ravenscroft., 1998) or microcontact printing of cell adhesion molecules such as polylycine (Jun et al., 2007) and laminin (Hammarback et al., 1985). However, long-term confinement of neuronal cultures to pre-determined geometry on a planar surface remains challenging due to detachment or degradation of surface adhesion molecules, surface masking by serum or cell secreted proteins, as well as cell migration and formation of axon bundles between cell clusters that distort the original pattern design (Ravenscroft., 1998, Branch et al., 2000). These short-term (i.e., <1 wk) cultures have limited capacity to generate VE-821 robust networks since cultures take weeks (>21 days) to fully develop mature axon connections (Dichter., 1978, Brewer et al., 1993). For these reasons, microfluidic structures such as microchannels (or tunnels) for the physical isolation of neuronal cells have been intensively investigated to generate long-term cultured neural networks (Claverol-Tinture and Pine., 2002, Bani-Yaghoub et al., 2005, Morin et al., 2005, Ravula et al., 2006, Dworak and Wheeler., 2009). While much progress has been made, the short distance of axons (500 m) in these small neural circuits are not ideal for axon tract accessibility. To establish functional connectivity of a neural circuit, two physically separated but axonally connected neuronal assemblies are expected to exhibit temporally related activities characteristic of their neuronal compositions. Those studies are routinely performed in live brain slices taking advantage of the well-established directionality of axon pathways. studies typically use multi-electrode arrays (MEAs) to record extracellular potential or current changes consequent of neuronal firing. These VE-821 multi-electrode recordings detect network activity changes and signal transmission, including action potential propagation (Dworak and Wheeler., 2009), network synchronization (Takayama et al., 2012) and conversation (Kanagasabapathi et al., 2011). Though useful for providing network-level analysis, MEA signals from cultured circuits lack characteristic patterns, and difficult to relate to biochemical and molecular events in single cells, such as channel protein deficits in axon injury (Iwata et al., 2004,Yuen et al., 2009). Recently, a long-length (2 mm) axon tract culture system (Tang-Schomer et al., 2010) was established as an model for studying diffuse axonal injury after brain injury. This model permits examination of powerful adjustments of axons (Tang-Schomer et al., 2010) and dendrites (Monnerie et al., 2010), such as for example mechanised injury-induced beading and undulations, that act like the pathological presentations of individual brains (Tang-Schomer et al., 2012). A youthful edition of compartmentalized civilizations, where axons grew as 2 mm-wide systems, was used to recognize sodium VE-821 route cleavage being a potential molecular system for human brain trauma-related useful deficits (Iwata et al., 2004, Yuen et al., 2009). Micropatterning methods were introduced to the model (Tang et al.,, 2005) to examine axon-tract particular adjustments in morphology (Tang-Schomer et al., 2010) aswell as channel proteins deficits (Tang-Schomer et SMARCB1 al, 2009). While guaranteeing, the prevailing model lacks VE-821 the capability to detect electrophysiological adjustments. In this scholarly study, we searched for to integrate the initial style feature of lengthy length axon tracts of our model using the features of electrophysiological and pharmacological manipulations right into a multi-functional gadget. To create the fabrication procedure, we try to integrate regular bioanalytical strategies that exist in neuroscience laboratories easily, so the gadget could be quickly modified for various other types of axon tract-associated CNS disorders. Here, we demonstrate an array of four paired two-node connections of 2 mm-long axon tracts that are integrated on planar MEAs. Local VE-821 perturbation of the circuit was exhibited with microfluidic application of live cell dyes as well as targeted electrical stimulation to individual nodes of the network. Functional connectivity was evaluated by local application of neurotransmitters to.