Immunohistochemical staining of wholemount major pelvic ganglia (MPG) for analysis of myelinated bladder afferents

This protocol describes immunohistochemical procedures applied to wholemount major pelvic ganglia (MPG) for the visualization of myelinated axons filled with cholera toxin subunit B (CTB) and/or neurons transfected with adeno-associated virus (AAV) expressing TdTomato. In this protocol, samples were obtained from rats in which CTB was microinjected into the bladder, and AAV-PHP.S was intravenously administered to preferentially and sparsely label peripheral neurons. In this context, neurofascin (paranode marker) identifies myelinated axons.