MP27-12 FUNCTIONAL ERECTOGENIC NERVE MAPPING OF THE ENTIRE RAT PROSTATE WITH OPTICAL CLEARING

Abstract

You have accessJournal of UrologyCME1 Apr 2023MP27-12 FUNCTIONAL ERECTOGENIC NERVE MAPPING OF THE ENTIRE RAT PROSTATE WITH OPTICAL CLEARING Selman Unal, Biljana Musicki, Ahmet Hoke, and Arthur Burnett Selman UnalSelman Unal More articles by this author , Biljana MusickiBiljana Musicki More articles by this author , Ahmet HokeAhmet Hoke More articles by this author , and Arthur BurnettArthur Burnett More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003255.12AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Erectile dysfunction is an important complication of radical prostatectomy (RP) due to cavernous nerve (CN) injury, which is in part a result of the unknown exact distribution of the CN branches (CNBs) on the prostate. We hypothesized that preserving functional pro-erectile CNBs promotes the success of retaining erectile function after RP. The aim of this study was to determine the functional erectogenic nerve distribution surrounding the entire prostate using a rat model. METHODS: Submaximal electrical stimulation of the CN was performed in male rats to induce penile erection by stimulating nNOS phosphorylation on Ser-1412 (functional nNOS). For localization of the P-nNOS (Ser-1412)-containing and nNOS-containing nerves, whole mount immunofluorescence staining was performed on the prostate, using ß-III-tubulin, a non-specific nerve marker, as control. Furthermore, the staining was combined with optical clearing with benzylalcohol/benzylbenzoate (BABB), which removes lipids and minimizes light scattering, allowing imaging of the tissue in situ. Because the BABB clearing technique has not been used previously for the CN or the whole prostate, we first modified it by optimizing stringency of lipid removal. RESULTS: We achieved optimum staining signal and prostate transparency by modification of BABB clearing technique to include shortening dehydration to prevent fluorescence quenching, followed by incubation with tert-butanol and BABB (1:2) at decreased acidity for 1.5 hour (Figure 1a). Microscopic examination localized nNOS and P-nNOS (Ser-1412)-containing cell bodies in the major pelvic ganglia, CN, and CNBs on the prostate. All nerves stained with nNOS/P-nNOS were also stained with ß-III-tubulin (Figure 1b, c). While there was no background noise in ß-III-tubulin staining, a minor signal of the nNOS and P-nNOS (Ser-1412) staining was observed in other tissues such as vascular smooth muscles. CONCLUSIONS: This study presents the first mapping of the pro-erectile nerves, in contrast to all nerves, surrounding the anatomically preserved whole rat prostate. This approach allows a dimensional shift from 2D to 3D histology of the whole intact prostate, providing further information about the distribution of pro-erectile CNBs in penile erection. Source of Funding: This study was supported by the Patrick C. Walsh Prostate Cancer Research Grant. Selman Unal was supported by The Scientific and Technological Research Council of Turkey (TÜBİTAK) for this study © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e366 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Selman Unal More articles by this author Biljana Musicki More articles by this author Ahmet Hoke More articles by this author Arthur Burnett More articles by this author Expand All Advertisement PDF downloadLoading ...