Muscarinic receptor function, density and G-protein coupling in the overactive diabetic rat bladder
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1. Bladder smooth muscle sensitivity to muscarinic agonists is increased in the overactive bladder. Treatment of rats with streptozotocin induces a diabetic state in which the bladder muscle is overactive and also supersensitive to muscarinic agonists. This study has examined bladder contraction, muscarinic receptor density and receptor/G-protein coupling in the streptozotocin-induced overactive bladder of the rat.
2. Diabetes was induced by a single intraperitoneal dose of streptozotocin. Seven days later contraction of isolated detrusor muscle strips was assessed in tissue bath experiments, while receptor density was assayed in saturation experiments with [3H]-QNB (quinuclidinyl benzilate, L-[benzilic-4,4¢-3H]) and receptor/G-protein coupling was determined in agonist displacement experiments with this radioligand.
3. Isolated detrusor strips from diabetic animals displayed an enhanced degree of spontaneous activity (0.060 ± 0.016 g mg)1, compared with 0.015 ± 0.004 g mg)1, P < 0.05). Carbachol produced contractile responses in tissues from both control and diabetic rats, but the diabetic tissues were more sensitive to this agonist, the pEC50 being 6.52 ± 0.17 compared with 5.93 ± 0.06 in controls (P < 0.001). Maximum responses to carbachol were similar in both groups of animals. The increase in carbachol potency was accompanied by a 40% increase in receptor density from 158 ± 5 to 221 ± 22 fmol mg)1 protein (P < 0.05), but this was not enough to fully account for the change in tissue sensitivity.
4. In the absence of GTP-c-S, carbachol displaced [3H]-QNB from two binding sites, the high affinity site (pKi ¼ 7.06 ± 0.26) which represents the receptors coupled to G-proteins made up 43.1 ± 5.9% of the total binding sites in control tissues and this value was similar (41.0 ± 4.0%) in the diabetic tissues (pKi ¼ 6.64 ± 0.31). In the presence of GTP-c-S, carbachol displaced [3H]-QNB from a single binding site which had a low-affinity, similar to the low-affinity site observed in the absence of GTP-c-S.
5. These data demonstrate that detrusor supersensitivity is observed after only 1 week of untreated diabetes in the rat. The overactivity is associated with an enhanced sensitivity to carbachol, which is partly explained by an increase in receptor density, but there appears to be no change in receptor/G-protein coupling.
© 2006 The Authors Journal compilation © 2006 Blackwell Publishing Ltd