Characterization of axons expressing the artemin receptor in the female rat urinary bladder: A comparison with other major neuronal populations

Artemin is a member of the glial cell line‐derived neurotrophic factor (GDNF) family that has been strongly implicated in development and regeneration of autonomic nerves and modulation of nociception. Whereas other members of this family (GDNF and neurturin) primarily target parasympathetic and nonpeptidergic sensory neurons, the artemin receptor (GFRα3) is expressed by sympathetic and peptidergic sensory neurons that are also the primary sites of action of nerve growth factor, a powerful modulator of bladder nerves. Many bladder sensory neurons express GFRα3 but it is not known if they represent a specific functional subclass. Therefore, our initial aim was to map the distribution of GFRα3‐immunoreactive (‐IR) axons in the female rat bladder, using cryostat sections and whole wall thickness preparations. We found that GFRα3‐IR axons innervated the detrusor, vasculature, and urothelium, but only part of this innervation was sensory. Many noradrenergic sympathetic axons innervating the vasculature were GFRα3‐IR, but the noradrenergic innervation of the detrusor was GFRα3‐negative. We also identified a prominent source of nonneuronal GFRα3‐IR that is likely to be glial. Further characterization of bladder nerves revealed specific structural features of chemically distinct classes of axon terminals, and a major autonomic source of axons labeled with neurofilament‐200, which is commonly used to identify myelinated sensory axons within organs. Intramural neurons were also characterized and quantified. Together, these studies reveal a diverse range of potential targets by which artemin could influence bladder function, nerve regeneration, and pain, and provide a strong microanatomical framework for understanding bladder physiology and pathophysiology. J. Comp. Neurol. 522:3900–3927, 2014. © 2014 Wiley Periodicals, Inc.