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IS MUSCARINIC RECEPTOR EXPRESSION ELEVATED IN URINARY BLADDER FROM WOMEN WITH RECURRENT URINARY TRACT INFECTIONS?
453
454
SUBUROTHELIAL MUSCARINIC RECEPTORS M1, M2, M3 IN PATIENTS WITH DETRUSOR OVERACTIVITY (DO) AND THE EFFECT OF BOTULINUM TOXIN TYPE A (BONT/A)
IS MUSCARINIC RECEPTOR EXPRESSION ELEVATED IN URINARY BLADDER FROM WOMEN WITH RECURRENT URINARY TRACT INFECTIONS?
Datta S.N.1, Roosen A.2, Popat R., Elneil S., Dasgupta P.4, Fowler C.J.1, Apostolidis A.1
0DQVᚏHOG.-1, Chandran J.2, Vaux K.J., Burcher E.2
1
1 University of Wollongong, Graduate School of Medicine, Wollongong, Australia, 2University of New South Wales, Dept. of Pharmacology, Sydney, Australia, Sydney Adventist Hospital, Dept. of Urology, Sydney, Australia
Institute of Neurology and National Hospital for Neurology and Neurosurgery, Dept. of Uro-Neurology, London, United Kingdom, 2Institute of Neurology, Dept. of Uro-Neurology, London, United Kingdom, National Hospital for Neurology and Neurosurgery, Dept. of Uro-Neurology, London, United Kingdom, 4 Guys’ and St. Thomas’ Hospitals and National Hospital for Neurology and Neurosurgery, Dept. of Urology, London, United Kingdom Introduction & Objectives: 0XVFDULQLFUHFHSWRUV00DQG0KDYHEHHQLGHQWLᚏHGERWKLQWKH detrusor and mucosa of the human bladder. It has been proposed that in overactive bladders increased release of Acetylcholine (ACh) by the urothelium acts on suburothelial and detrusor muscarinic UHFHSWRUVUHVXOWLQJLQ'2,QFUHDVHGSUHVHQFHRIVXEXURWKHOLDO0DQG0UHFHSWRUVKDVEHHQVKRZQ in human idiopathic DO, whereas M1 receptors appear to play a facilitatory role in the release of ACh and noradrenaline in spinalised animal bladders. It is thought the urothelium, suburothelial nerves and P\RᚏEUREODVWVIXQFWLRQDVDXQLWWRSURYLGHEODGGHUVHQVRU\LQIRUPDWLRQ%R17$NQRZQWREORFNWKH UHOHDVHRI$&KKDVDOVREHHQVKRZQWRDᚎHFWKXPDQVXEXURWKHOLDOVHQVRU\SDWKZD\VLQSDWLHQWVZLWK LQWUDFWDEOH'2:HLQYHVWLJDWHGWKHWUHDWPHQWHᚎHFWRILQWUDGHWUXVRU%R17$RQVXEXURWKHOLDO00 DQG0UHFHSWRUVLQ'2SDWLHQWVDQGLQFRPSDULVRQZLWKFRQWUROV Material & Methods: )ROORZLQJ(WKLFV&RPPLWWHHDSSURYDOᚐH[LEOHF\VWRVFRS\EODGGHUELRSVLHVZHUH obtained from 24 patients with either neurogenic or idiopathic DO (NDO/IDO) at baseline and at 4 and 16 weeks after successful BoNT/A treatment, and from 6 controls with asymptomatic microscopic KDHPDWXULD 6SHFLPHQV ZHUH LPPXQRVWDLQHG RYHUQLJKW XVLQJ VSHFLᚏF DQWLERGLHV WR PXVFDULQLF UHFHSWRUVVXEW\SHV,PPXQRUHDFWLYLW\ ,5 ZDVTXDQWLᚏHGZLWKLPDJHDQDO\VLVDQGH[SUHVVHGDV SHUFHQWDJHSHUKLJKSRZHUᚏHOG7KH0DQQ:KLWQH\DQG.UXVNDO:DOOLVWHVWVZHUHXVHGIRUVWDWLVWLFDO comparisons accordingly.
Introduction & Objectives: A history of recurrent urinary tract infection (rUTI) is associated with an increased risk of developing bladder overactivity (Kjolhede et al 1997, Acta Obstet Gynecol 6FDQG 7KHDLPRIWKLVVWXG\ZDVWRFRPSDUHWKHOHYHORIH[SUHVVLRQRI0DQG0 muscarinic receptors in bladder detrusor and mucosa in women with a history of recurrent UTI, with that in bladder from asymptomatic women. Material & Methods: %ODGGHUELRSVLHVIURPFRQWUROV \HDUV DQGSDWLHQWVZLWKDKLVWRU\RI recurrent UTI (21-87 years) were collected into RNA LaterTM, with approval from human ethics committees, and dissected into detrusor and mucosa. RNA was extracted and expression of P51$IRU0DQG0PXVFDULQLFUHFHSWRUVDQGIRUWKHLQWHUQDOFRQWURO*$3'+ZDVGHWHUPLQHG XVLQJ TXDQWLWDWLYH FRPSHWLWLYH 573&5 DV GHVFULEHG SUHYLRXVO\ 0DQVᚏHOG HW DO %U - Pharmacol, 144, 1089-99). Muscarinic receptor mRNA expression was normalised to expression RI*$3'+LQWKHVDPHVDPSOH5HVXOWVDUHSUHVHQWHGDVPHGLDQVZLWK FRQᚏGHQFHOLPLWV
Results: Suburothelial muscarinic receptor IR was observed in cells with bipolar processes and ᚏEUHOLNHVWUXFWXUHV$GHFUHDVHLQWRWDOVXEXURWKHOLDO0,5ZDVQRWHGLQ'2ELRSVLHVFRPSDUHGWR FRQWUROV S )ROORZLQJ%R17$WKHUHZDVLQFUHDVHDQGಫQRUPDOLVDWLRQಬRI0,5 S DQG DWDQGZHHNVUHVSHFWLYHO\ (TXDOO\VLJQLᚏFDQWSRVW%R17$LQFUHDVHVZHUHVHHQLQ0,5 S DQGDWDQGZHHNVUHVSHFWLYHO\ &RQWUROELRSVLHVZHUHLQDGHTXDWHIRUEDVHOLQH FRPSDULVRQVRI0,5'HVSLWHDGHFUHDVHLQ0,5LQEDVHOLQH'2VSHFLPHQV S FRPSDUHG WRFRQWUROV%R17$WUHDWPHQWKDGQRHᚎHFWRQWRWDOVXEXURWKHOLDO0,5
Results: 7KHUHZDVQRGLᚎHUHQFHLQH[SUHVVLRQRIPXVFDULQLF0UHFHSWRUP51$LQGHWUXVRU IURPUHFXUUHQW87,SDWLHQWVFRPSDUHGWRWKDWRIDJHPDWFKHGFRQWUROV >@SJQJ DQG >@ SJ QJ UHVSHFWLYHO\ Q 6LPLODUO\ QR FKDQJH LQ GHWUXVRU 0 P51$ H[SUHVVLRQFRXOGEHGHPRQVWUDWHG UHFXUUHQW87,>@SJQJFRQWURO>@ pg/ 100ng, n=11-12). GAPDH expression in control and recurrent UTI samples was equal in WKHGHWUXVRU&RPSDUHGZLWKFRQWUROPXFRVD >ದ@SJQJ51$Q 3 GAPDH expression was consistently lower in the mucosa from recurrent UTI patients (0.05 >@ SJ QJ 51$ Q LQGLFDWLQJ WKDW P51$ LQ PXFRVD IURP WKHVH SDWLHQWV ZDV degraded. However, this did not correspond with degradation of detrusor mRNA in the same ELRSV\ >@SJQJ51$ :HKDYHSUHYLRXVO\GHPRQVWUDWHGP51$GHJUDGDWLRQ in the mucosa from biopsies of women with interstitial cystitis, a condition also associated with PXFRVDOLQᚐDPPDWLRQ /LXHWDO1HXURXURO8URG\Q
Conclusions: Decreased levels of suburothelial muscarinic receptors in DO patients may be in accord ZLWKSUHYLRXVᚏQGLQJVRIUHGXFHGGHWUXVRUPXVFDULQLFUHFHSWRUOHYHOVLQ'2WKDWZHUHDFFRPSDQLHG by functional hyperexcitability. Post-BoNT/A increase in suburothelial muscarinic receptor levels LV XQOLNHO\ WR EH GXH WR D GLUHFW SUROLIHUDWLYH HᚎHFW DQG FRXOG EH FRPSHQVDWRU\ WR WKH QHHGV RI D ORZIUHTXHQF\IXQFWLRQLQJEODGGHUZLWKUHGXFHGUHOHDVHRI$&KVXSSRUWLQJDQHXURSODVWLFHᚎHFWRI %R17$RQEODGGHUDᚎHUHQWSDWKZD\V7KHUHVXOWVVKRXOGEHDOVRDQDO\VHGLQWKHOLJKWRISRVVLEOH detrusor receptor changes.
Conclusions: In conclusion, the apparent association in females of recurrent UTI with detrusor RYHUDFWLYLW\LVXQOLNHO\WREHGXHWRFKDQJHVLQH[SUHVVLRQRIPXVFDULQLF0RU0UHFHSWRUVLQWKH GHWUXVRUPXVFOH:HKDYHSUHYLRXVO\GHPRQVWUDWHGDGHFUHDVHLQ0H[SUHVVLRQLQZRPHQZLWK LGLRSDWKLFGHWUXVRURYHUDFWLYLW\ 0DQVᚏHOGHWDO%-8,QW DQGZHK\SRWKHVLVHG that a similar decrease in muscarinic mRNA expression in bladder mucosa from women with recurrent UTI would be seen, but this could not be examined because of degradation of mucosal RNA.
455
P28 SEXUAL FUNCTION: EJACULATION AND FEMALE FUNCTION Thursday, 27 March, 12.15-13.45, Blue Hall 2
THE NON NEURONAL CHOLINERGIC SYSTEM OF THE HUMAN UROTHELIUM- IS THERE A CONTRIBUTION TO THE PATHOGENESIS OF DETRUSOR OVERACTIVITY?
BRAIN OXYTOCIN RECEPTOR BLOCKADE INHIBITS PHARMACOLOGICALLY INDUCED SEXUAL RESPONSES IN ANAESTHETISED MALE RATS
Bschleipfer T.1, Schukowski K.1, Weidner W.1, Grando S.A.2, Schwantes U., Kummer W.4, Lips K.S.4
Clement P.1, Peeters M.1, Bernabe J.1, Laurin M.1, Denys P.2, Giuliano F.2 1
1
Justus-Liebig-University Giessen, Dept. of Urology and Pediatric Urology, Giessen, Germany, 2 Uc Davis Medical Group, Dept. of Dermatology, Sacramento, United States of America, Fa. 3ᚐHJHU *PE+ 'HSW RI 0HGLFDO 6FLHQFH &OLQLFDO 5HVHDUFK %DPEHUJ *HUPDQ\ 4JustusLiebig-University Giessen, Institute for Anatomy and Cell Biology, Giessen, Germany Introduction & Objectives: In prior studies we could demonstrate a non neurological cholinergic system (NNCS) inside the human urothelium of female individuals. Its components seem to be able to synthesize and release acetylcholine (ACh). The expression and distribution of muscarinic and nicotinic cholinergic receptors could be described as well. This study aimed WRLQYHVWLJDWHGLᚎHUHQFHVEHWZHHQWKHIHPDOHDQGWKHPDOHXURWKHOLXPUHIHUULQJWRWKLVV\VWHP )XUWKHUPRUHLWVKRXOGEHFODULᚏHGLIWKH11&6LVLQYROYHGLQWKHSDWKRJHQHVLVRIGHWUXVRURYHU activity. Material & Methods: :HH[DPLQHGWHQIHPDOHDQGVL[PDOHSDWLHQWVZKRGLGQRWVXᚎHUIURP detrusor over activity (DO), urgency or urge incontinence. A third group encompassed eight male patients showing DO stemming from subvesical obstruction (BOO) proved by means of F\VWRPHWU\ $FWXDO XULQDU\ LQIHFWLRQ RU QHXURORJLFDO GHᚏFLHQFLHV DᚎHFWLQJ WKH XULQDU\ EODGGHU ZHUHH[FOXGHGLQDOOLQGLYLGXDOV'XULQJHQGRVFRSLFVXUJHU\VXSHUᚏFLDOVSHFLPHQVZHUHWDNHQ out of the bottom of the bladder gathering the urothelium only. The samples were examined by real-time RT-PCR and product size was validated by gel electrophoresis. We examined and compared the appearance of muscarinic receptor subtypes M1R to M5R, choline acetyltransferase (ChAT), choline transporter-1 (CHT1), vesicular acetylcholine transporter 9$&K7 RUJDQLF FDWLRQ WUDQVSRUWHUV 2&7 WR 2&7 DQG QLFRWLQLF DFHW\OFKROLQH UHFHSWRU subunits alpha7, -9 and -10. Results: ,QIHPDOHSDWLHQWV573&5GHWHFWHGWKHWUDQVFULSWVRI&+72&7DQG2&7ZKLOH it failed detecting ChAT, VAChT and OCT2. Referring to the receptors, expression of all types of muscarinic receptor subtypes (M1R to M5R) and nicotinic receptor subunits (alpha7, -9 and -10) was observed. The investigation of the male specimens showed similar results with no GLᚎHUHQFHWRWKHIHPDOHV\VWHP&RPSDULQJEODGGHUVDPSOHVIURPQRQREVWUXFWLYHPDOHSDWLHQWV ZLWKWKRVHRISDWLHQWVVXᚎHULQJIURPEODGGHURXWOHWREVWUXFWLRQUHVXOWLQJLQGHWUXVRURYHUDFWLYLW\ ZHIRXQG&+72&7DQGDVZHOODVWKHPXVFDULQLFDQGQLFRWLQLFUHFHSWRUVDWWKHVDPH expression level while transcripts of ChAT, VAChT and OCT2 could also not be detected. Conclusions: This study proves the similarity of the non neuronal cholinergic system of human urothelium between both genders. This study gives also rise to the assumption that the non neuronal cholinergic system might not be involved in the pathogenesis of detrusor over activity.
Eur Urol Suppl 2008;7(3):184
2
456
Pelvipharm Laboratories, Dept. of Neuropharmacology, Gif-sur-Yvette, France, Poincare Hospital, Dept. of Physical Medicine and Rehabilitation, Garches, France
Introduction & Objectives: The present study was undertaken to clarify the role of brain, spinal, and peripheral oxytocin (OT) receptors in the control of ejaculation and HUHFWLRQ )RU WKLV SXUSRVH ZH H[SORUHG LQ DQDHVWKHWLVHG UDWV WKH HᚎHFWV RI DQ 27 DQWDJRQLVW 27 DQWDJ GHOLYHUHG WKURXJK GLᚎHUHQW URXWHV RQ VH[XDO UHVSRQVHV HOLFLWHG by intracerebroventricular (i.c.v.) 7-hydroxy-2-(di-N-propylamino)tetralin (7-OH-DPAT), a GRSDPLQH'SUHIHUHQWLDODJRQLVW Material & Methods: A cannula for i.c.v. injection was stereo tactically implanted to sexually naive adult male Wistar rats anaesthetised with urethane. Seminal vesicle pressure and bulbospongiosus muscle (BS) electromyogram were recorded as physiological markers of, respectively, emission and expulsion phases of ejaculation. Intracavernosal pressure was measured as physiological marker of erection. The peptidergic selective OT antag (d(CH2)51,Tyr(Me)2,Orn8)-Oxytocin) was delivered i.v. (0.5 and 1 µg/kg, n=10 each), i.c.v. DQGwJQ HDFK RULQWUDWKHFDOO\ wJLWDWWKHthWKRUDFLF 7 or 6th OXPEDU / VHJPHQW Q HDFK SULRU WR LFY 2+'3$7 6H[XDO UHVSRQVHV ZHUHUHFRUGHGRYHUPLQIROORZLQJ2+'3$7DGPLQLVWUDWLRQ Results: Whatever the dose tested i.v., the OT antag did not impair 7-OH-DPAT-induced sexual responses occurring in form of coordinated increases in seminal vesicle and intracavernosal pressures and rhythmic BS contractions sometime accompanied by expulsion of seminal material (i.e. ejaculation). When delivered i.c.v., the OT antag doseGHSHQGHQWO\ LQKLELWHG 2+'3$7LQGXFHG VH[XDO UHVSRQVHV WKH PD[LPDO LFY GRVH tested (0.1 µg) resulting in abolition of ejaculation. When delivered i.t. (0.1 µg) at L6 but QRWDW7WKH27DQWDJVLJQLᚏFDQWO\UHGXFHGWKHGXUDWLRQRI%6FRQWUDFWLRQVDQGWKH occurrence of ejaculation without impairing erectile response. Conclusions: From these results it is concluded that, in the 7-OH-DPAT model, (i) brain OT receptors mediate ejaculation and erection, and (ii) L6 spinal OT receptors have a modulating role on ejaculation but not on erection whereas (iii) peripheral OT receptors are not involved in either ejaculation or erection. In addition the present data support the existence of functional relationships between dopaminergic and oxytocinergic pathways in the central control of sexual responses.
454
SUBUROTHELIAL MUSCARINIC RECEPTORS M1, M2, M3 IN PATIENTS WITH DETRUSOR OVERACTIVITY (DO) AND THE EFFECT OF BOTULINUM TOXIN TYPE A (BONT/A)
IS MUSCARINIC RECEPTOR EXPRESSION ELEVATED IN URINARY BLADDER FROM WOMEN WITH RECURRENT URINARY TRACT INFECTIONS?
Datta S.N.1, Roosen A.2, Popat R., Elneil S., Dasgupta P.4, Fowler C.J.1, Apostolidis A.1
0DQVᚏHOG.-1, Chandran J.2, Vaux K.J., Burcher E.2
1
1 University of Wollongong, Graduate School of Medicine, Wollongong, Australia, 2University of New South Wales, Dept. of Pharmacology, Sydney, Australia, Sydney Adventist Hospital, Dept. of Urology, Sydney, Australia
Institute of Neurology and National Hospital for Neurology and Neurosurgery, Dept. of Uro-Neurology, London, United Kingdom, 2Institute of Neurology, Dept. of Uro-Neurology, London, United Kingdom, National Hospital for Neurology and Neurosurgery, Dept. of Uro-Neurology, London, United Kingdom, 4 Guys’ and St. Thomas’ Hospitals and National Hospital for Neurology and Neurosurgery, Dept. of Urology, London, United Kingdom Introduction & Objectives: 0XVFDULQLFUHFHSWRUV00DQG0KDYHEHHQLGHQWLᚏHGERWKLQWKH detrusor and mucosa of the human bladder. It has been proposed that in overactive bladders increased release of Acetylcholine (ACh) by the urothelium acts on suburothelial and detrusor muscarinic UHFHSWRUVUHVXOWLQJLQ'2,QFUHDVHGSUHVHQFHRIVXEXURWKHOLDO0DQG0UHFHSWRUVKDVEHHQVKRZQ in human idiopathic DO, whereas M1 receptors appear to play a facilitatory role in the release of ACh and noradrenaline in spinalised animal bladders. It is thought the urothelium, suburothelial nerves and P\RᚏEUREODVWVIXQFWLRQDVDXQLWWRSURYLGHEODGGHUVHQVRU\LQIRUPDWLRQ%R17$NQRZQWREORFNWKH UHOHDVHRI$&KKDVDOVREHHQVKRZQWRDᚎHFWKXPDQVXEXURWKHOLDOVHQVRU\SDWKZD\VLQSDWLHQWVZLWK LQWUDFWDEOH'2:HLQYHVWLJDWHGWKHWUHDWPHQWHᚎHFWRILQWUDGHWUXVRU%R17$RQVXEXURWKHOLDO00 DQG0UHFHSWRUVLQ'2SDWLHQWVDQGLQFRPSDULVRQZLWKFRQWUROV Material & Methods: )ROORZLQJ(WKLFV&RPPLWWHHDSSURYDOᚐH[LEOHF\VWRVFRS\EODGGHUELRSVLHVZHUH obtained from 24 patients with either neurogenic or idiopathic DO (NDO/IDO) at baseline and at 4 and 16 weeks after successful BoNT/A treatment, and from 6 controls with asymptomatic microscopic KDHPDWXULD 6SHFLPHQV ZHUH LPPXQRVWDLQHG RYHUQLJKW XVLQJ VSHFLᚏF DQWLERGLHV WR PXVFDULQLF UHFHSWRUVVXEW\SHV,PPXQRUHDFWLYLW\ ,5 ZDVTXDQWLᚏHGZLWKLPDJHDQDO\VLVDQGH[SUHVVHGDV SHUFHQWDJHSHUKLJKSRZHUᚏHOG7KH0DQQ:KLWQH\DQG.UXVNDO:DOOLVWHVWVZHUHXVHGIRUVWDWLVWLFDO comparisons accordingly.
Introduction & Objectives: A history of recurrent urinary tract infection (rUTI) is associated with an increased risk of developing bladder overactivity (Kjolhede et al 1997, Acta Obstet Gynecol 6FDQG 7KHDLPRIWKLVVWXG\ZDVWRFRPSDUHWKHOHYHORIH[SUHVVLRQRI0DQG0 muscarinic receptors in bladder detrusor and mucosa in women with a history of recurrent UTI, with that in bladder from asymptomatic women. Material & Methods: %ODGGHUELRSVLHVIURPFRQWUROV \HDUV DQGSDWLHQWVZLWKDKLVWRU\RI recurrent UTI (21-87 years) were collected into RNA LaterTM, with approval from human ethics committees, and dissected into detrusor and mucosa. RNA was extracted and expression of P51$IRU0DQG0PXVFDULQLFUHFHSWRUVDQGIRUWKHLQWHUQDOFRQWURO*$3'+ZDVGHWHUPLQHG XVLQJ TXDQWLWDWLYH FRPSHWLWLYH 573&5 DV GHVFULEHG SUHYLRXVO\ 0DQVᚏHOG HW DO %U - Pharmacol, 144, 1089-99). Muscarinic receptor mRNA expression was normalised to expression RI*$3'+LQWKHVDPHVDPSOH5HVXOWVDUHSUHVHQWHGDVPHGLDQVZLWK FRQᚏGHQFHOLPLWV
Results: Suburothelial muscarinic receptor IR was observed in cells with bipolar processes and ᚏEUHOLNHVWUXFWXUHV$GHFUHDVHLQWRWDOVXEXURWKHOLDO0,5ZDVQRWHGLQ'2ELRSVLHVFRPSDUHGWR FRQWUROV S )ROORZLQJ%R17$WKHUHZDVLQFUHDVHDQGಫQRUPDOLVDWLRQಬRI0,5 S DQG DWDQGZHHNVUHVSHFWLYHO\ (TXDOO\VLJQLᚏFDQWSRVW%R17$LQFUHDVHVZHUHVHHQLQ0,5 S DQGDWDQGZHHNVUHVSHFWLYHO\ &RQWUROELRSVLHVZHUHLQDGHTXDWHIRUEDVHOLQH FRPSDULVRQVRI0,5'HVSLWHDGHFUHDVHLQ0,5LQEDVHOLQH'2VSHFLPHQV S FRPSDUHG WRFRQWUROV%R17$WUHDWPHQWKDGQRHᚎHFWRQWRWDOVXEXURWKHOLDO0,5
Results: 7KHUHZDVQRGLᚎHUHQFHLQH[SUHVVLRQRIPXVFDULQLF0UHFHSWRUP51$LQGHWUXVRU IURPUHFXUUHQW87,SDWLHQWVFRPSDUHGWRWKDWRIDJHPDWFKHGFRQWUROV >@SJQJ DQG >@ SJ QJ UHVSHFWLYHO\ Q 6LPLODUO\ QR FKDQJH LQ GHWUXVRU 0 P51$ H[SUHVVLRQFRXOGEHGHPRQVWUDWHG UHFXUUHQW87,>@SJQJFRQWURO>@ pg/ 100ng, n=11-12). GAPDH expression in control and recurrent UTI samples was equal in WKHGHWUXVRU&RPSDUHGZLWKFRQWUROPXFRVD >ದ@SJQJ51$Q 3 GAPDH expression was consistently lower in the mucosa from recurrent UTI patients (0.05 >@ SJ QJ 51$ Q LQGLFDWLQJ WKDW P51$ LQ PXFRVD IURP WKHVH SDWLHQWV ZDV degraded. However, this did not correspond with degradation of detrusor mRNA in the same ELRSV\ >@SJQJ51$ :HKDYHSUHYLRXVO\GHPRQVWUDWHGP51$GHJUDGDWLRQ in the mucosa from biopsies of women with interstitial cystitis, a condition also associated with PXFRVDOLQᚐDPPDWLRQ /LXHWDO1HXURXURO8URG\Q
Conclusions: Decreased levels of suburothelial muscarinic receptors in DO patients may be in accord ZLWKSUHYLRXVᚏQGLQJVRIUHGXFHGGHWUXVRUPXVFDULQLFUHFHSWRUOHYHOVLQ'2WKDWZHUHDFFRPSDQLHG by functional hyperexcitability. Post-BoNT/A increase in suburothelial muscarinic receptor levels LV XQOLNHO\ WR EH GXH WR D GLUHFW SUROLIHUDWLYH HᚎHFW DQG FRXOG EH FRPSHQVDWRU\ WR WKH QHHGV RI D ORZIUHTXHQF\IXQFWLRQLQJEODGGHUZLWKUHGXFHGUHOHDVHRI$&KVXSSRUWLQJDQHXURSODVWLFHᚎHFWRI %R17$RQEODGGHUDᚎHUHQWSDWKZD\V7KHUHVXOWVVKRXOGEHDOVRDQDO\VHGLQWKHOLJKWRISRVVLEOH detrusor receptor changes.
Conclusions: In conclusion, the apparent association in females of recurrent UTI with detrusor RYHUDFWLYLW\LVXQOLNHO\WREHGXHWRFKDQJHVLQH[SUHVVLRQRIPXVFDULQLF0RU0UHFHSWRUVLQWKH GHWUXVRUPXVFOH:HKDYHSUHYLRXVO\GHPRQVWUDWHGDGHFUHDVHLQ0H[SUHVVLRQLQZRPHQZLWK LGLRSDWKLFGHWUXVRURYHUDFWLYLW\ 0DQVᚏHOGHWDO%-8,QW DQGZHK\SRWKHVLVHG that a similar decrease in muscarinic mRNA expression in bladder mucosa from women with recurrent UTI would be seen, but this could not be examined because of degradation of mucosal RNA.
455
P28 SEXUAL FUNCTION: EJACULATION AND FEMALE FUNCTION Thursday, 27 March, 12.15-13.45, Blue Hall 2
THE NON NEURONAL CHOLINERGIC SYSTEM OF THE HUMAN UROTHELIUM- IS THERE A CONTRIBUTION TO THE PATHOGENESIS OF DETRUSOR OVERACTIVITY?
BRAIN OXYTOCIN RECEPTOR BLOCKADE INHIBITS PHARMACOLOGICALLY INDUCED SEXUAL RESPONSES IN ANAESTHETISED MALE RATS
Bschleipfer T.1, Schukowski K.1, Weidner W.1, Grando S.A.2, Schwantes U., Kummer W.4, Lips K.S.4
Clement P.1, Peeters M.1, Bernabe J.1, Laurin M.1, Denys P.2, Giuliano F.2 1
1
Justus-Liebig-University Giessen, Dept. of Urology and Pediatric Urology, Giessen, Germany, 2 Uc Davis Medical Group, Dept. of Dermatology, Sacramento, United States of America, Fa. 3ᚐHJHU *PE+ 'HSW RI 0HGLFDO 6FLHQFH &OLQLFDO 5HVHDUFK %DPEHUJ *HUPDQ\ 4JustusLiebig-University Giessen, Institute for Anatomy and Cell Biology, Giessen, Germany Introduction & Objectives: In prior studies we could demonstrate a non neurological cholinergic system (NNCS) inside the human urothelium of female individuals. Its components seem to be able to synthesize and release acetylcholine (ACh). The expression and distribution of muscarinic and nicotinic cholinergic receptors could be described as well. This study aimed WRLQYHVWLJDWHGLᚎHUHQFHVEHWZHHQWKHIHPDOHDQGWKHPDOHXURWKHOLXPUHIHUULQJWRWKLVV\VWHP )XUWKHUPRUHLWVKRXOGEHFODULᚏHGLIWKH11&6LVLQYROYHGLQWKHSDWKRJHQHVLVRIGHWUXVRURYHU activity. Material & Methods: :HH[DPLQHGWHQIHPDOHDQGVL[PDOHSDWLHQWVZKRGLGQRWVXᚎHUIURP detrusor over activity (DO), urgency or urge incontinence. A third group encompassed eight male patients showing DO stemming from subvesical obstruction (BOO) proved by means of F\VWRPHWU\ $FWXDO XULQDU\ LQIHFWLRQ RU QHXURORJLFDO GHᚏFLHQFLHV DᚎHFWLQJ WKH XULQDU\ EODGGHU ZHUHH[FOXGHGLQDOOLQGLYLGXDOV'XULQJHQGRVFRSLFVXUJHU\VXSHUᚏFLDOVSHFLPHQVZHUHWDNHQ out of the bottom of the bladder gathering the urothelium only. The samples were examined by real-time RT-PCR and product size was validated by gel electrophoresis. We examined and compared the appearance of muscarinic receptor subtypes M1R to M5R, choline acetyltransferase (ChAT), choline transporter-1 (CHT1), vesicular acetylcholine transporter 9$&K7 RUJDQLF FDWLRQ WUDQVSRUWHUV 2&7 WR 2&7 DQG QLFRWLQLF DFHW\OFKROLQH UHFHSWRU subunits alpha7, -9 and -10. Results: ,QIHPDOHSDWLHQWV573&5GHWHFWHGWKHWUDQVFULSWVRI&+72&7DQG2&7ZKLOH it failed detecting ChAT, VAChT and OCT2. Referring to the receptors, expression of all types of muscarinic receptor subtypes (M1R to M5R) and nicotinic receptor subunits (alpha7, -9 and -10) was observed. The investigation of the male specimens showed similar results with no GLᚎHUHQFHWRWKHIHPDOHV\VWHP&RPSDULQJEODGGHUVDPSOHVIURPQRQREVWUXFWLYHPDOHSDWLHQWV ZLWKWKRVHRISDWLHQWVVXᚎHULQJIURPEODGGHURXWOHWREVWUXFWLRQUHVXOWLQJLQGHWUXVRURYHUDFWLYLW\ ZHIRXQG&+72&7DQGDVZHOODVWKHPXVFDULQLFDQGQLFRWLQLFUHFHSWRUVDWWKHVDPH expression level while transcripts of ChAT, VAChT and OCT2 could also not be detected. Conclusions: This study proves the similarity of the non neuronal cholinergic system of human urothelium between both genders. This study gives also rise to the assumption that the non neuronal cholinergic system might not be involved in the pathogenesis of detrusor over activity.
Eur Urol Suppl 2008;7(3):184
2
456
Pelvipharm Laboratories, Dept. of Neuropharmacology, Gif-sur-Yvette, France, Poincare Hospital, Dept. of Physical Medicine and Rehabilitation, Garches, France
Introduction & Objectives: The present study was undertaken to clarify the role of brain, spinal, and peripheral oxytocin (OT) receptors in the control of ejaculation and HUHFWLRQ )RU WKLV SXUSRVH ZH H[SORUHG LQ DQDHVWKHWLVHG UDWV WKH HᚎHFWV RI DQ 27 DQWDJRQLVW 27 DQWDJ GHOLYHUHG WKURXJK GLᚎHUHQW URXWHV RQ VH[XDO UHVSRQVHV HOLFLWHG by intracerebroventricular (i.c.v.) 7-hydroxy-2-(di-N-propylamino)tetralin (7-OH-DPAT), a GRSDPLQH'SUHIHUHQWLDODJRQLVW Material & Methods: A cannula for i.c.v. injection was stereo tactically implanted to sexually naive adult male Wistar rats anaesthetised with urethane. Seminal vesicle pressure and bulbospongiosus muscle (BS) electromyogram were recorded as physiological markers of, respectively, emission and expulsion phases of ejaculation. Intracavernosal pressure was measured as physiological marker of erection. The peptidergic selective OT antag (d(CH2)51,Tyr(Me)2,Orn8)-Oxytocin) was delivered i.v. (0.5 and 1 µg/kg, n=10 each), i.c.v. DQGwJQ HDFK RULQWUDWKHFDOO\ wJLWDWWKHthWKRUDFLF 7 or 6th OXPEDU / VHJPHQW Q HDFK SULRU WR LFY 2+'3$7 6H[XDO UHVSRQVHV ZHUHUHFRUGHGRYHUPLQIROORZLQJ2+'3$7DGPLQLVWUDWLRQ Results: Whatever the dose tested i.v., the OT antag did not impair 7-OH-DPAT-induced sexual responses occurring in form of coordinated increases in seminal vesicle and intracavernosal pressures and rhythmic BS contractions sometime accompanied by expulsion of seminal material (i.e. ejaculation). When delivered i.c.v., the OT antag doseGHSHQGHQWO\ LQKLELWHG 2+'3$7LQGXFHG VH[XDO UHVSRQVHV WKH PD[LPDO LFY GRVH tested (0.1 µg) resulting in abolition of ejaculation. When delivered i.t. (0.1 µg) at L6 but QRWDW7WKH27DQWDJVLJQLᚏFDQWO\UHGXFHGWKHGXUDWLRQRI%6FRQWUDFWLRQVDQGWKH occurrence of ejaculation without impairing erectile response. Conclusions: From these results it is concluded that, in the 7-OH-DPAT model, (i) brain OT receptors mediate ejaculation and erection, and (ii) L6 spinal OT receptors have a modulating role on ejaculation but not on erection whereas (iii) peripheral OT receptors are not involved in either ejaculation or erection. In addition the present data support the existence of functional relationships between dopaminergic and oxytocinergic pathways in the central control of sexual responses.