Abstract
The ability of peripheral nervous system neurons to extend long, axon-like neurites in vitro makes them ideally suited for studies on mechanisms of axon survival and degeneration. In this chapter, we describe how to prepare explant cultures of sympathetic neurons of the superior cervical ganglion (SCG). We also describe how to induce and assess axon degeneration with an injury or a chemical insult.
Highlights
Much of the current knowledge on mechanisms of axon degeneration comes from studies in primary neuronal cultures of sympathetic superior cervical ganglion (SCG) and sensory dorsal root ganglion (DRG) neurons
Neuronal cell bodies are located within the ganglion, with long axon-like neurites (5–6 mm long at 7 days in vitro, DIV) radially extending away from it; this hugely simplifies experiments of axon degeneration following an injury since all neurites distal to the site of injury are transected (Fig. 1)
We describe two methods to induce neurite degeneration: (1) the injury model, where degeneration is initiated by a physical transection of the neurites, a process known as Wallerian degeneration, and (2) chemically induced degeneration, where neurite death is caused by the administration of a toxic compound to uninjured neurons
Summary
Axon loss is an early feature of several neurodegenerative disorders and leads to compromised neuronal function. Understanding how axons die is important to identify therapeutic targets to delay or halt the progression of these pathologies [1]. Much of the current knowledge on mechanisms of axon degeneration comes from studies in primary neuronal cultures of sympathetic superior cervical ganglion (SCG) and sensory dorsal root ganglion (DRG) neurons
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