Axon tracing and node segmentation of myelinated bladder afferents in the pelvic nerve v1

This protocol describes method for tracing myelinated axons labelled with the adeno-associated virus, AAV-PHP.S and neurofascin immunohistochemistry (to identify paranodes) in pelvic nerve of rat, using Neurolucida 360. This tracing method can be applied to various types of peripheral nerves. In rats, AAV-PHP.S has a high tropism for myelinated but not unmyelinated afferents. Cholera toxin subunit B (CTB) microinjected into the bladder was used to identify bladder afferents in the pelvic nerve. This protocol does not include details of the immunohistochemistry or procedures involving animals (viral labelling and CTB tracing). Other antibodies relevant to myelinated axons can be used instead of neurofascin.