Methods in Neuronal Growth Cone Biology

Axon guidance is a crucial component of normal nervous system development. Errors in axon guidance can result in disorders such as autism and schizophrenia. In order to understand the pathogenesis of these disorders, we need to understand the fundamental mechanisms of growth cone motility, which ultimately direct axon guidance. In this chapter, we describe protocols and methods that our laboratory uses to study normal growth cone physiology and biochemistry. We describe the “pipette” turning assay [Lohof AM, Quillan M, Dan Y, Poo MM, J Neurosci 12(4): 1253–1261, 1992] that we have adapted for primary cultures of rodent dorsal root ganglia. We also describe methods for calcium imaging of growth cones and provide data to demonstrate the absolute requirement of TRP channels for growth cone navigation. These techniques provide researchers with considerable scope and flexibility in experimental design and can be adapted to provide high-throughput biochemical data for the elucidation of mechanisms governing the action of guidance molecules.