Millipore Technical Publications |
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Poster: High Throughput Screening To Identify Novel Molecular Pathways Involved In Neurite Outgrowth | ||
| Lit No: | ps1039en00 | |
| Year: | 2008 | |
Abstract
An abundance of data has been generated with the completion of the genome project which provides a comprehensive view of kinases and the molecular regulators of the cell. However, very little is known about the mechanism of these regulators and their relationship to molecular pathways responsible for the development of complex three-dimensional tissues. To acquire a better understanding, some labs have already employed a combination of functional genomics (loss of function) screen with morphological analysis in explant cultures. Here, to discover novel kinase regulated molecular pathways in neurite outgrowth we have used Neurite Outgrowth Assay for High Content Screening to screen for gain or loss of neuritis in neuronal cells exposed to siRNA kinome. Furthermore, neurite outgrowth was quantified using Neurite Outgrowth Quantification Kit. To that end, we have shown that neurite outgrowth in N1E-115 neuroblastoma cells was significantly stimulated in Millicell® inserts coated with the ECM protein laminin compared to BSA controls (*p<0.05; n=5). Furthermore, application of 75μM nocodazole, a microtubuledestabilizing drug, reduced the formation of neurites. We then quantitated the effect of knocking down 18 different kinases on neurite outgrowth. The characterization of neurite formation, maturation and collapse/resorption is an area of intense interest, since these cellular processes are essential for interconnection of neuronal cell bodies. Therefore, major efforts in central nervous system drug discovery research are focused on the identification of compounds that affect neurite outgrowth. In summary, our results demonstrate an unbiased approach in understanding the relationship of molecular pathways and their impact on morphological parameters in cellular systems.Click the PDF to view the entire poster.
