The microtubule-associated protein Tau is primarily expressed in neurons and plays an integral role in the regulation of multiple functions within the axon. In the adult brain, the six Tau isoforms are expressed allowing for a complex system of control. Despite Tau's central role, the mechanisms by which Tau acts are not fully understood. We have used single-molecule total internal reflection fluorescence (TIRF) microscopy and the methods described in this chapter to further our knowledge of Tau's behavior and function. We have demonstrated that Tau's dynamic binding behavior allows for regulation of motor protein motility and microtubule dynamics in an isoform-specific manner. The continued use and refinement of the single-molecule techniques detailed here can only further our knowledge of Tau and other proteins integral to the maintenance of axonal transport.
Keywords: Kinesin; Microscopy; Microtubules; Single-molecule; Tau; Total internal reflection fluorescence (TIRF).
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