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POSTER PRESENTATION Open Access Distinct ageing- and androgen-dependent effects on cGMP signalling proteins in three male rat urogenital organs (bladder, prostate, epididymis) Dieter Müller 1* , Michail S Davidoff 2 , Ralf Middendorff 1 From 5th International Conference on cGMP: Generators, Effectors and Therapeutic Implications Halle, Germany. 24-26 June 2011 Background Aging of the male reproductive system leads to changes in endocrine signalling and is frequently associated with the emergence of bothersome conditions owing to pros- tate hyperplasia and bladder dysfunctions. Recent findings suggest that urogenital organs are promising targets for therapeutic interventions with inhibitors of the cGMP- degrading phosphodiesterase 5 (PDE5) [1]. However, the cGMP signalling system in these tissues and possible age- related alterations are only partially characterized. This study investigated key proteins of cGMP pathways in bladder, prostate and epididymis of young (3 months) and old (23-24 months) Wistar rats. To address andro- gen-dependent effects, we (i) used an experimental model, where injection of ethane dimethane sulphonate (EDS) into adult rats evokes a temporary elimination of the testosterone-producing Leydig cells [2] and (ii) exam- ined effects induced by castration and testosterone supplementation. Methods and results Western blot analyses aimed to characterize the levels of PDE5, soluble (sGC) and particulate (GC-A, GC-B) guany- lyl cyclases, endothelial nitric oxide synthase (eNOS) and cGMP-dependent protein kinase I (cGKI) in membrane and cytosolic protein fractions. Data revealed a strikingly low abundance of all these proteins in prostate. In con- trast, the bladder was distinguished by exceptionally high levels of PDE5, cGKI and eNOS, each resembling those in lung tissue. The epididymis was characterized by a predo- minance of particulate (in particular GC-B) versus soluble (sGC) guanylyl cyclases and lowest PDE5 tissue concentrations. As evidenced also by membrane guanylyl cyclase assays, ageing did not affect the three (GC-A, GC-B, sGC) cGMP- generating enzymes in all organs. However, and supported by PDE activity studies, we recognized a pronounced age- related decrease (by nearly 50%) of PDE5 in bladder. Membrane-associated (but not cytosolic) cGKI concentra- tions were markedly reduced in the aged epididymis. On the other hand, prostatic cGKI increased with ageing. Androgen withdrawal during temporary Leydig cell elimi- nation induced a massive (>12-fold) upregulation of cGKI in prostate but not other (penis, epididymis) androgen- dependent organs. Castration elicited an equally organ- selective upregulation of cGKI in prostate. Neither EDS treatment nor castration evoked increases in other kinases (PKA, Akt, PI3K, GSK-3) examined. Testosterone supple- mentation reduced the increased cGKI levels in prostates of castrated rats in a dose-dependent manner. Conclusion The findings may have significance for ageing-associated pathologies in the male lower urinary tract and identify cGKI as an androgen-sensitive signalling protein in prostate. Author details 1 Institute of Anatomy and Cell Biology, University of Giessen, Germany. 2 Institute of Clinical, Chemistry, University of Hamburg, Germany. Published: 1 August 2011 References 1. Sandner P, Neuser D, Bischoff E: Erectile dysfunction and lower urinary tract. Handb Exp Pharmacol 2009, 191:507-531. * Correspondence: [email protected] 1 Institute of Anatomy and Cell Biology, University of Giessen, Germany Full list of author information is available at the end of the article Müller et al. BMC Pharmacology 2011, 11(Suppl 1):P51 http://www.biomedcentral.com/1471-2210/11/S1/P51 © 2011 Müller et al; licensee BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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POSTER PRESENTATION Open Access

Distinct ageing- and androgen-dependent effectson cGMP signalling proteins in three male raturogenital organs (bladder, prostate, epididymis)Dieter Müller1*, Michail S Davidoff2, Ralf Middendorff1

From 5th International Conference on cGMP: Generators, Effectors and Therapeutic ImplicationsHalle, Germany. 24-26 June 2011

BackgroundAging of the male reproductive system leads to changesin endocrine signalling and is frequently associated withthe emergence of bothersome conditions owing to pros-tate hyperplasia and bladder dysfunctions. Recent findingssuggest that urogenital organs are promising targets fortherapeutic interventions with inhibitors of the cGMP-degrading phosphodiesterase 5 (PDE5) [1]. However, thecGMP signalling system in these tissues and possible age-related alterations are only partially characterized. Thisstudy investigated key proteins of cGMP pathways inbladder, prostate and epididymis of young (3 months)and old (23-24 months) Wistar rats. To address andro-gen-dependent effects, we (i) used an experimentalmodel, where injection of ethane dimethane sulphonate(EDS) into adult rats evokes a temporary elimination ofthe testosterone-producing Leydig cells [2] and (ii) exam-ined effects induced by castration and testosteronesupplementation.

Methods and resultsWestern blot analyses aimed to characterize the levels ofPDE5, soluble (sGC) and particulate (GC-A, GC-B) guany-lyl cyclases, endothelial nitric oxide synthase (eNOS) andcGMP-dependent protein kinase I (cGKI) in membraneand cytosolic protein fractions. Data revealed a strikinglylow abundance of all these proteins in prostate. In con-trast, the bladder was distinguished by exceptionally highlevels of PDE5, cGKI and eNOS, each resembling those inlung tissue. The epididymis was characterized by a predo-minance of particulate (in particular GC-B) versus soluble

(sGC) guanylyl cyclases and lowest PDE5 tissueconcentrations.As evidenced also by membrane guanylyl cyclase assays,

ageing did not affect the three (GC-A, GC-B, sGC) cGMP-generating enzymes in all organs. However, and supportedby PDE activity studies, we recognized a pronounced age-related decrease (by nearly 50%) of PDE5 in bladder.Membrane-associated (but not cytosolic) cGKI concentra-tions were markedly reduced in the aged epididymis. Onthe other hand, prostatic cGKI increased with ageing.Androgen withdrawal during temporary Leydig cell elimi-nation induced a massive (>12-fold) upregulation of cGKIin prostate but not other (penis, epididymis) androgen-dependent organs. Castration elicited an equally organ-selective upregulation of cGKI in prostate. Neither EDStreatment nor castration evoked increases in other kinases(PKA, Akt, PI3K, GSK-3) examined. Testosterone supple-mentation reduced the increased cGKI levels in prostatesof castrated rats in a dose-dependent manner.

ConclusionThe findings may have significance for ageing-associatedpathologies in the male lower urinary tract and identifycGKI as an androgen-sensitive signalling protein inprostate.

Author details1Institute of Anatomy and Cell Biology, University of Giessen, Germany.2Institute of Clinical, Chemistry, University of Hamburg, Germany.

Published: 1 August 2011

References1. Sandner P, Neuser D, Bischoff E: Erectile dysfunction and lower urinary

tract. Handb Exp Pharmacol 2009, 191:507-531.* Correspondence: [email protected] of Anatomy and Cell Biology, University of Giessen, GermanyFull list of author information is available at the end of the article

Müller et al. BMC Pharmacology 2011, 11(Suppl 1):P51http://www.biomedcentral.com/1471-2210/11/S1/P51

© 2011 Müller et al; licensee BioMed Central Ltd. This is an open access article distributed under the terms of the Creative CommonsAttribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction inany medium, provided the original work is properly cited.

2. Davidoff MS, Middendorff M, Enikolopov G, Riethmacher D, Holstein AF,Müller D: Progenitor cells of the testosterone-producing Leydig cellsrevealed. J Cell Biol 2004, 167:935-944.

doi:10.1186/1471-2210-11-S1-P51Cite this article as: Müller et al.: Distinct ageing- and androgen-dependent effects on cGMP signalling proteins in three male raturogenital organs (bladder, prostate, epididymis). BMC Pharmacology2011 11(Suppl 1):P51.

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