2004 Fiscal Year Final Research Report Summary
The role of autonomic nerve receptor subtypes and G-proteins for the cause of overactive bladder
| Project/Area Number |
14571516
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Urology
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| Research Institution | Dokkyo University School of Medicine |
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Principal Investigator |
YAMANISHI Tomonori Dokkyo University, School of Medicine, Department of Urology, Associate professor, 医学部, 助教授 (90220425)
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| Co-Investigator(Kenkyū-buntansha) |
KAMAI Takao Dokkyo University, School of Medicine, Lecturer, 医学部, 講師 (80316562)
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| Project Period (FY) |
2002 – 2004
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| Keywords | bladder / smooth muscle / muscarinic / adrenergic / Rho / Rho-kinase / carbachol / β-adrenoceptor |
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| Research Abstract |
The role of muscarinic receptor subtypes in mediating contraction of detrusor muscles : The affinities of muscarinic antagonists against carbachol responses were calculated in normal and cAMP-elevated tissues (contraction with KCl and relaxation with isoprenaline) using longitudinal and transverse strips of urethra. The pig urethra appears to possess predominantly M_2 muscarinic receptors. Contraction of the normal urethra appears to be predominantly mediated by M_2 and M_3-receptors in circular muscle, but by M_3-receptors in longitudinal muscle. Following cAMP-elevation, a contribution to contraction of M_2-receptors appeared to be demonstrated in both tissues, but the involvement of the M_2-receptors appeared greater in the circular muscle.
The role of β_3-Adrenoceptors in mediating relaxation of bladder and urethral smooth muscle : of the pig and human. : Cell membranes were prepared from detrusor and urethral muscle of the pig and competition experiments with [^3H]-dihydroalprenolo
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l(DHA), a non-selective β-adrenoceptor antagonist was used as a specific radioligand to determine the presence of β-adrenoceptor subtypes. In functional experiments, isolated detrusor muscle strips were used to determine the potency of agonists and the affinity of antagonists. β_3-adrenoceptars are the predominant β-adrenoceptor subtype present in the pig bladder and urethra. Functionally, β-adrenoceptor mediated responses of this tissue are mediated via both the β_2- and β_3-adrenoceptor subtypes. β_3-adrenoceptors appear to be more involved in relaxation of bladder body in the pig and human, whereas β_2-adrenoceptors appear to be more involved in trigonal and urethral smooth muscle of the pig.
Rho/Rho kinase is predominantly present in the urothelium controlling the contraction of the human urinary bladder in vitro : Samples of detrusor muscle from the dome region of the human urinary bladder were obtained from patients undergoing total cystectomy for bladder cancer. The mRNA and protein levels of Rho-kinase were analyzed using the relative multiplex RT-PCR and Western blotting. Immunohistochemistry, using the same specific antibodies as antibodies for Western blotting, was performed to support the data obtained by Western blotting. Cumulative concentration-response curves(CRCs) to carbachol were obtained, and were then equilibrated for 30 mm with Krebs solution containing the 10μM of Y 27632 or vehicle (time control). On the relative multiplex RT-PCR,RhoA and Rho kinase were more abundant in the urothelium than in the smooth muscle. Western blotting also demonstrated significantly greater expression of Rho kinase protein expression in the urothelium than in the smooth muscle (p<0.0001 and p<0.0001, respectively). Urothelial cells showed moderate to strong reaction for Rho kinase antibodies, and smooth muscle cells showed weak to moderate reaction on immunohistochemistrical study. Rho and Rho kinase appear to be predominantly present in the urothelium. The maximum response to carbachol was decreased in the presence of urothelium. The Rho/Rho kinase pathway appears to be involved in the contraction of human bladder smooth muscle in the presence of urothelium. Less
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