Mapping of functional erectogenic nerves on the rat prostate

Background Preservation of erectogenic nerves during radical prostatectomy (RP) is hampered by limited understanding of their precise localization, due to their complex, intertwined paths, and the inherent individual variations across patients. Because erection utilizes a subset of cavernous nerves (CNs) that in response to sexual stimuli reveal phosphorylation of neuronal nitric oxide synthase (nNOS) on its stimulatory site Ser-1412, we hypothesized that delineation of nerves containing phosphorylated (P)-nNOS on Ser-1412 would establish the location of functional erectogenic nerves within the periprostatic region.Aim To identify the distribution and quantity of functional erection-relevant ([P-nNOS]-containing) nerves in the periprostatic area and discriminate them among the CNs distribution. We further evaluated whether functional communication exists between contralateral CNs.Methods Young adult male Sprague-Dawley rats underwent electrical stimulation of the CNs to induce penile erection via phosphorylation of nNOS on Ser-1412 (6 V for 2 min, n = 6). No stimulation group served as control (n = 6). The prostate and adjacent structures were collected and processed for whole-mount double-staining with TuJ1 antibody (a pan-axonal marker) and P-nNOS (n = 3 for stimulation, n = 3 for no stimulation), or total nNOS and P-nNOS (n = 3 for stimulation, n = 3 for no stimulation), followed by modified optical clearing and microscopic examination. Nerve quantification was done by systematic counting.Outcomes Location and quantification of functional erectogenic nerves.Results In the male rat, we obtained a map of P-nNOS-containing nerves in the periprostatic area, which are relevant for penile erection. Only 17.5% of all nerves, and only 28.4% of the total nNOS-containing nerves in the periprostatic region are functionally erectogenic nerves. Furthermore, there is no functional innervation between contralateral (stimulated and non-stimulated) CNs.Clinical Implications This basic science study is expected to provide a foundation for subsequent studies at the human level. Understanding the erection-relevant nerve distribution in the periprostatic area is expected to advance nerve-sparing RP at the human level to improve sexual function outcomes.Strengths and Limitations This is the first study to describe and quantitate a subset of functional erection-relevant (P-nNOS-containing) nerves in the periprostatic area. Our study differs from previous studies where nerves containing total nNOS were localized without specifying which nerves produce erection. However, because this study comprised a relatively small number of rats, further studies with a bigger sample size or other model animals are warranted.Conclusion Only a subset of nerve fibers in the periprostatic region represent functional erectogenic nerves, characterized by the expression of P-nNOS (Ser-1412).