| Complex structures at both the gross and cellular levels endow the nervous system with its distinct function: the rapid processing of information via elaborate patterns of synaptic connectivity. Various arrangements of numerous growth factors, guidance cues, and other molecules influence both the interconnectivity of neurons and the subcellular makeup of individual neurons. This dissertation describes experiments that used an engineering-based approach to study the mechanisms that regulate neuronal behavior, both in small networks and at the subcellular level. The first half of this report shows that high-resolution protein patterns and cellular positioning systems may eventually be used to develop simple, reliable neuronal networks. Specifically, a PDMS stencil was used to restrict cellular positions at plating to specific regions on a micropatterned culturing substrate with the goal of regulating neurite outgrowth and neuronal network formation. Also, a variety of protein patterns were tested to determine their effects on the direction of axonal extension. Next, to look at subcellular events during axon outgrowth, the second half of the dissertation utilized a PDMS microdevice to isolate purified axonal mRNA transcripts for microarray analysis. The list of axonal mRNAs generated will facilitate further studies of protein synthesis within axons, which is now a recognized process in normal axon physiology. |
