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Structural Autonomic Evaluation in Chronic Pelvic Pain Syndromes
Abstracts / Autonomic Neuroscience: Basic and Clinical 192 (2015) 56–141
P1.3 ATP evokes cholinergic contraction in the healthy, but not inflamed, intact rat urinary bladder J. Stenqvist, M. Winder, M. Johnsson, G. Tobin, P. Aronsson Dept. Pharmacology, University of Gothenburg, Sweden Background: Over the past years increasing evidence suggesting an altered role of purinergic signaling during pathological conditions has been presented. Furthermore, there have been speculations regarding a possible link between purinergic and cholinergic functional mechanisms. In order to further investigate this, an “intact (whole) bladder” in vitro-setup has presently been employed. Aim: To elucidate the role of the urothelium and the functional link between purinergic and cholinergic signaling in the healthy and inflamed intact rat urinary bladder. Methods: Intact urinary bladders were excised from anesthetized rats; controls or cyclophosphamide-treated (100 mg/kg 60 h before the experiment to induce cystitis). Two catheters were inserted into the bladder for administration of substances and outflow, respectively. Collagenase I was used to denude the urothelium. Functional studies were conducted in an organ bath system. Agonists and antagonists could be administered both into the surrounding Krebs solution and intravesically (via the catheters). Results and Conclusions: When instilling ATP into the intact bladder, a contraction was observed (5.8 ± 0.9 mN; 1 mM; n = 19) which, interestingly, was significantly decreased in the presence of atropine (1 μM; 2.2 ± 0.6 mN, p b 0.01; n = 11). This difference was not observed neither in inflamed (2.8 ± 0.4 and 2.4 ± 0.1 mN; n = 3-8), nor in urothelium denuded bladders (3.0 ± 1.3 and 3.3 ±2.0 mN; n = 4-8). This preliminary study shows the delicate interactions between the two transmitter systems, namely that ATP partly induces detrusor muscle contractions indirectly by stimulating the release of acetylcholine. Also, the urothelium seems to be involved in this cholinergic response, which appears to be diminished during inflammation.
doi:10.1016/j.autneu.2015.07.012
P1.4 Cholinergic activation of brain stem neurons increases intravesical pressure mediated by vasopressin release in female rats E.M. Cafarchioa, R.L. Almeidaa, C.A. Ogiharaa, L.C. Valdoa, M.C.B. Luzb, F.L.A. Fonsecab, J.S. Souzac, R.M.B. Macielc, G. Giannoccoc, M.A. Satoa a Dept. Morfology and Physiology, Faculdade de Medicina do ABC, FMABC, Brazil b Clinical Analysis Laboratory, FMABC, Brazil c Dept. Biological Sciences, Federal University of Sao Paulo, Brazil Background: Previous findings of our laboratory have shown that injection of the cholinergic agonist carbachol into the 4th brain ventricle (4th V) increases intravesical pressure (IP) with peak response at 30 minutes after injection. Aim: We investigated if carbachol injection into the 4th V induces vasopressin (AVP) or oxytocin (OT) release and the existence of AVP and OT receptors in the urinary bladder (UB). Methods: Female Wistar rats (~250 g, N = 6/group) implanted with guide cannulas into the 4th V five days prior to the experiments were anesthetized with 2% isoflurane in 100% O2, and carbachol (4 nmol/μL) or saline (1 μL) was injected into the 4th V. Thirty minutes after injection, a blood sample was collected and plasma AVP and OT was determined by ELISA kit (Cayman Chemical). In a different group of rats, gene expression in the UB for AVP1A, AVP1B, AVP2, OT recepμtors and
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cyclophilin (housekeeping gene) were investigated by qPCR. Data are as mean ± SE and were submitted to Student t-test (p b 0.05). Results: Carbachol induced plasma AVP release (3051 ± 85 pg/mL) compared to saline into the 4th V (2628 ± 276 pg/mL). No difference was observed in plasma OT in rats which received carbachol (532 ± 33 pg/ mL) or saline (508 ± 38 pg/mL). All subytpes of AVP and OT receptors showed their genes expressed in the UB. Conclusion: Cholinergic activation of brain stem neurons by carbachol induces plasma AVP release. The increase in IP evoked by carbachol is likely due to AVP release which binds to AVP receptors in the urinary bladder. Financial support: FAPESP (grant# 2013/04550-5) and NEPAS.
doi:10.1016/j.autneu.2015.07.013
P1.5 Structural Autonomic Evaluation in Chronic Pelvic Pain Syndromes G. Chelimsky, P. McCabe, T. Chelimsky Medical College of Wisconsin, USA Background: Functional abnormalities of the autonomic nervous system (ANS) often occur in chronic pain, including chronic pelvic pain (CPP). Hypothesis: abnormal ANS innervation of the bladder underlies bladder pain syndrome (BPS). We therefore compared ANS testing in pa-tients with 2 types of CPP: BPS and myofascial pelvic pain (MPP), and healthy control subjects (HC). Methods: In this IRB approved protocol, with 39 HC, 36 BPS, 14 MPP and 41 BPS + MPP sub-jects underwent one cardiac parasympathetic test, [deep breathing (DB)], two cardiac and vas-cular autonomic tests [valsalva maneuver and tilt table test (HUT)], and post-ganglionic sudo-motor sympathetic test that evaluated for autonomic neuropathy (AN). A validated composite autonomic laboratory score was applied. Results: Cardiac response to DB did not differ among groups. HUT was more frequently abnormal in CPP than HC, (36/91, 40% vs 4/39, 10%, p b 0.001), most frequent diagnosis was orthostatic intolerance. Physiologic HUT diagnoses such as orthostatic hypotension, postural tachycardia syndrome and syncope occurred rarely. AN was more frequent in CPP (28/91, 31% vs 5/39, 10% p = 0.01). CPP groups did not differ from one another in either of these abnormalities. Discussion: Our preliminary study on 14 BPS subjects and 15 HC found no structural autonomic abnormality in CPP. This final report based on 130 subjects confirms absence of autonomic cardiovascular abnormalities. Symptoms in the upright position (orthostatic intolerance) on tilt may suggest central sensitization. The increased frequency of AN in almost a third of subjects with CPP needs further investigation. Funding Source: ICEPAC project funded by NIH-NIDDK R01DK083538.
doi:10.1016/j.autneu.2015.07.014
P1.6 Receiver operating characteristic analysis of sphincter electromyography and post-void residuals for multiple system atrophy T. Yamamotoa, R. Sakakibarad, T. Uchiyamaa,e, M. Fuseb, M. Yanagisawab, Y. Kogac, S. Kuwabaraa a Department of Neurology, Chiba University Hospital, Chiba, Japan b Department of Urology, Chiba University Hospital, Chiba, Japan c Department of Clinical laboratory, Chiba University Hospital, Chiba, Japan
P1.3 ATP evokes cholinergic contraction in the healthy, but not inflamed, intact rat urinary bladder J. Stenqvist, M. Winder, M. Johnsson, G. Tobin, P. Aronsson Dept. Pharmacology, University of Gothenburg, Sweden Background: Over the past years increasing evidence suggesting an altered role of purinergic signaling during pathological conditions has been presented. Furthermore, there have been speculations regarding a possible link between purinergic and cholinergic functional mechanisms. In order to further investigate this, an “intact (whole) bladder” in vitro-setup has presently been employed. Aim: To elucidate the role of the urothelium and the functional link between purinergic and cholinergic signaling in the healthy and inflamed intact rat urinary bladder. Methods: Intact urinary bladders were excised from anesthetized rats; controls or cyclophosphamide-treated (100 mg/kg 60 h before the experiment to induce cystitis). Two catheters were inserted into the bladder for administration of substances and outflow, respectively. Collagenase I was used to denude the urothelium. Functional studies were conducted in an organ bath system. Agonists and antagonists could be administered both into the surrounding Krebs solution and intravesically (via the catheters). Results and Conclusions: When instilling ATP into the intact bladder, a contraction was observed (5.8 ± 0.9 mN; 1 mM; n = 19) which, interestingly, was significantly decreased in the presence of atropine (1 μM; 2.2 ± 0.6 mN, p b 0.01; n = 11). This difference was not observed neither in inflamed (2.8 ± 0.4 and 2.4 ± 0.1 mN; n = 3-8), nor in urothelium denuded bladders (3.0 ± 1.3 and 3.3 ±2.0 mN; n = 4-8). This preliminary study shows the delicate interactions between the two transmitter systems, namely that ATP partly induces detrusor muscle contractions indirectly by stimulating the release of acetylcholine. Also, the urothelium seems to be involved in this cholinergic response, which appears to be diminished during inflammation.
doi:10.1016/j.autneu.2015.07.012
P1.4 Cholinergic activation of brain stem neurons increases intravesical pressure mediated by vasopressin release in female rats E.M. Cafarchioa, R.L. Almeidaa, C.A. Ogiharaa, L.C. Valdoa, M.C.B. Luzb, F.L.A. Fonsecab, J.S. Souzac, R.M.B. Macielc, G. Giannoccoc, M.A. Satoa a Dept. Morfology and Physiology, Faculdade de Medicina do ABC, FMABC, Brazil b Clinical Analysis Laboratory, FMABC, Brazil c Dept. Biological Sciences, Federal University of Sao Paulo, Brazil Background: Previous findings of our laboratory have shown that injection of the cholinergic agonist carbachol into the 4th brain ventricle (4th V) increases intravesical pressure (IP) with peak response at 30 minutes after injection. Aim: We investigated if carbachol injection into the 4th V induces vasopressin (AVP) or oxytocin (OT) release and the existence of AVP and OT receptors in the urinary bladder (UB). Methods: Female Wistar rats (~250 g, N = 6/group) implanted with guide cannulas into the 4th V five days prior to the experiments were anesthetized with 2% isoflurane in 100% O2, and carbachol (4 nmol/μL) or saline (1 μL) was injected into the 4th V. Thirty minutes after injection, a blood sample was collected and plasma AVP and OT was determined by ELISA kit (Cayman Chemical). In a different group of rats, gene expression in the UB for AVP1A, AVP1B, AVP2, OT recepμtors and
57
cyclophilin (housekeeping gene) were investigated by qPCR. Data are as mean ± SE and were submitted to Student t-test (p b 0.05). Results: Carbachol induced plasma AVP release (3051 ± 85 pg/mL) compared to saline into the 4th V (2628 ± 276 pg/mL). No difference was observed in plasma OT in rats which received carbachol (532 ± 33 pg/ mL) or saline (508 ± 38 pg/mL). All subytpes of AVP and OT receptors showed their genes expressed in the UB. Conclusion: Cholinergic activation of brain stem neurons by carbachol induces plasma AVP release. The increase in IP evoked by carbachol is likely due to AVP release which binds to AVP receptors in the urinary bladder. Financial support: FAPESP (grant# 2013/04550-5) and NEPAS.
doi:10.1016/j.autneu.2015.07.013
P1.5 Structural Autonomic Evaluation in Chronic Pelvic Pain Syndromes G. Chelimsky, P. McCabe, T. Chelimsky Medical College of Wisconsin, USA Background: Functional abnormalities of the autonomic nervous system (ANS) often occur in chronic pain, including chronic pelvic pain (CPP). Hypothesis: abnormal ANS innervation of the bladder underlies bladder pain syndrome (BPS). We therefore compared ANS testing in pa-tients with 2 types of CPP: BPS and myofascial pelvic pain (MPP), and healthy control subjects (HC). Methods: In this IRB approved protocol, with 39 HC, 36 BPS, 14 MPP and 41 BPS + MPP sub-jects underwent one cardiac parasympathetic test, [deep breathing (DB)], two cardiac and vas-cular autonomic tests [valsalva maneuver and tilt table test (HUT)], and post-ganglionic sudo-motor sympathetic test that evaluated for autonomic neuropathy (AN). A validated composite autonomic laboratory score was applied. Results: Cardiac response to DB did not differ among groups. HUT was more frequently abnormal in CPP than HC, (36/91, 40% vs 4/39, 10%, p b 0.001), most frequent diagnosis was orthostatic intolerance. Physiologic HUT diagnoses such as orthostatic hypotension, postural tachycardia syndrome and syncope occurred rarely. AN was more frequent in CPP (28/91, 31% vs 5/39, 10% p = 0.01). CPP groups did not differ from one another in either of these abnormalities. Discussion: Our preliminary study on 14 BPS subjects and 15 HC found no structural autonomic abnormality in CPP. This final report based on 130 subjects confirms absence of autonomic cardiovascular abnormalities. Symptoms in the upright position (orthostatic intolerance) on tilt may suggest central sensitization. The increased frequency of AN in almost a third of subjects with CPP needs further investigation. Funding Source: ICEPAC project funded by NIH-NIDDK R01DK083538.
doi:10.1016/j.autneu.2015.07.014
P1.6 Receiver operating characteristic analysis of sphincter electromyography and post-void residuals for multiple system atrophy T. Yamamotoa, R. Sakakibarad, T. Uchiyamaa,e, M. Fuseb, M. Yanagisawab, Y. Kogac, S. Kuwabaraa a Department of Neurology, Chiba University Hospital, Chiba, Japan b Department of Urology, Chiba University Hospital, Chiba, Japan c Department of Clinical laboratory, Chiba University Hospital, Chiba, Japan