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Objective: We previously found that mirabegron exerts a relaxant effect in the presence of the β 3 -adrenoceptor antagonist SR58894A during carbachol-induced contraction in human and pig detrusor. The aim of this study was to explore the possible mechanism underlying the relaxant effects of mirabegron using detrusor smooth muscle., Methods: Human tissue was obtained from urinary bladders of patients undergoing radical cystectomy at Kyushu University and Harasanshin Hospital. Pig tissue was obtained from an abattoir. Tension force (organ bath experiments) was measured in intact or permeabilised (α-toxin or β-escin) detrusor smooth muscle strips. The contribution of cAMP-dependent signaling and the inhibition of Ca 2+ sensitization to the relaxant effects of mirabegron were characterized using 1 μM SR58894A, 100 μM SQ22536 (an adenylyl cyclase inhibitor), 10 μM H-89 (a protein kinase [PK] A inhibitor), 10 μM Y-27632 (a selective Rho kinase inhibitor), and 10 μM GF-109203X (a selective PKC inhibitor)., Results: 30 μM Mirabegron impaired carbachol (0.03-1 μM)-induced contraction in human detrusor smooth muscle. SR58894A only partially attenuated the relaxant effects of mirabegron in human and pig detrusor strips precontracted with 1 μM carbachol. In α-toxin-permeabilized detrusor strips, tension force at 1 μM [Ca 2+ ] i was decreased by mirabegron in a concentration-dependent manner. The relaxant effect of mirabegron was only slightly attenuated by H-89 and not significantly affected by SQ22536. Y-27632 potentiated the relaxation response to mirabegron, but attenuated responses to cAMP; GF-109203X had little effect. Mirabegron but not cAMP had a notable relaxant effect in the pig detrusor smooth muscle permeabilized with β-escin., Conclusions: Mirabegron-induced relaxation of pig and human detrusor smooth muscle occurs via both a β 3 -adrenoceptor/cAMP-dependent and -independent pathway., (© 2019 John Wiley & Sons Australia, Ltd.)

Manganese is a neurotoxin causing manganism, a Parkinson-like clinical disorder. Manganese has been shown to interfere with dopaminergic neurotransmission, but the neurotoxic mechanism involved is not fully resolved. In the bivalve mollusc Crassostrea virginica also known as the eastern oyster, beating rates of lateral cilia of the gill are controlled by dopaminergic-serotonergic innervation originating from their cerebral and visceral ganglia. Terminal release of dopamine activates D2-like receptors on these gill cells inhibiting adenylyl cyclase and slowing cilia beating rates. In C. virginica, manganese treatment disrupts this dopaminergic innervation of the gill, preventing the normal cilio-inhibitory response of lateral cells to dopamine. In this study an adenylyl cyclase activator (forskolin) and two different inhibitors (MDL-12,330A and SQ 22,536) were used to determine if manganese had any effects on the adenylyl cyclase step of the dopamine D2 receptor signal transduction pathway. The results showed that neither the adenylyl cyclase activator nor the inhibitors were affected by manganese in the control of lateral ciliary activity. This suggests that in C. virginica the mechanism of manganese toxicity on the dopaminergic control of lateral ciliary activity is targeting an early step in the D2R signal transduction pathway, which may involve interference with D2 receptor activation or alternatively some other downstream signaling activity that does not affect adenylyl cyclase., (Copyright © 2018 Elsevier Inc. All rights reserved.)

For territorial animals, establishment of status-dependent dominance order is essential to maintain social stability. In agonistic encounters of the crayfish Procambarus clarkii, a difference of body length of 3-7% is enough for larger animals to become dominant. Despite a physical disadvantage, small winners of the first pairings were more likely to win subsequent conflicts with larger inexperienced animals. In contrast, the losers of the first pairings rarely won subsequent conflicts with smaller naive animals. Such experiences of previous winning or losing affected agonistic outcomes for a long period. The winner effects lasted more than 2 weeks and the loser effect lasted about 10 days. Injection of 5HT1 receptor antagonist into the dominant animals 15-30 min after establishment of dominance order blocked the formation of the winner effects. In contrast, injection of adrenergic-like octopamine receptor antagonist into subordinate animals blocked the formation of the loser. 5HT1 receptors are negatively coupled to adenylyl cyclase and adrenergic-like octopamine receptors are positively coupled. Consistent with this, dominant animals failed to show the winner effect when injected with pCPT-cAMP, a cAMP analogue, and subordinate animals failed to show a loser effect when injected with adenylyl cyclase inhibitor SQ 22536. These results suggest that an increase and decrease of cAMP concentration is essential in mediating loser and winner effects, respectively. Furthermore, formation of the loser effect was blocked by injection of protein kinase A (PKA) inhibitor H89, suggesting long-term memory of the loser effect is dependent on the cAMP-PKA signalling pathway., (© 2016 Federation of European Neuroscience Societies and John Wiley & Sons Ltd.)

Background and Purpose: Polymorphisms of the μ opioid receptor (MOPr) may contribute to the variation in responses to opioid drugs in clinical and unregulated situations. The A6V variant of MOPr (MOPr-A6V) is present in up to 20% of individuals in some populations, and may be associated with heightened susceptibility to drug abuse. There are no functional studies examining the acute signalling of MOPr-A6V in vitro, so we investigated potential functional differences between MOPr and MOPr-A6V at several signalling pathways using structurally distinct opioid ligands., Experimental Approach: CHO and AtT-20 cells stably expressing MOPr and MOPr-A6V were used. AC inhibition and ERK1/2 phosphorylation were assayed in CHO cells; K channel activation was assayed in AtT-20 cells., Key Results: Buprenorphine did not inhibit AC or stimulate ERK1/2 phosphorylation in CHO cells expressing MOPr-A6V, but buprenorphine activation of K channels in AtT-20 cells was preserved. [D-Ala2, N-MePhe4, Gly-ol]-enkephalin, morphine and β-endorphin inhibition of AC was significantly reduced via MOPr-A6V, as was signalling of all opioids to ERK1/2. However, there was little effect of the A6V variant on K channel activation., Conclusions and Implications: Signalling to AC and ERK via the mutant MOPr-A6V was decreased for many opioids, including the clinically significant drugs morphine, buprenorphine and fentanyl, as well endogenous opioids. The MOPr-A6V variant is common and this compromised signalling may affect individual responses to opioid therapy, while the possible disruption of the endogenous opioid system may contribute to susceptibility to substance abuse., (© 2014 The British Pharmacological Society.)