Structural determinants at the M2 muscarinic receptor modulate the RGS4-GIRK response to pilocarpine by impairment of the receptor voltage sensitivity.

Membrane potential controls the response of the M2 muscarinic receptor to its ligands. Membrane hyperpolarization increases response to the full agonist acetylcholine (ACh) while decreasing response to the partial agonist pilocarpine. We previously have demonstrated that the regulator of G-protein signaling (RGS) 4 protein discriminates between the voltage-dependent responses of ACh and pilocarpine; however, the underlying mechanism remains unclear. Here we show that RGS4 is involved in the voltage-dependent behavior of the M2 muscarinic receptor-mediated signaling in response to pilocarpine. Additionally we revealed structural determinants on the M2 muscarinic receptor underlying the voltage-dependent response. By electrophysiological recording in Xenopus oocytes expressing M2 muscarinic receptor and G-protein-gated inwardly rectifying K+ channels, we quantified voltage-dependent desensitization of pilocarpine-induced current in the presence or absence of RGS4. Hyperpolarization-induced desensitization of the current required for RGS4, also depended on pilocarpine concentration. Mutations of charged residues in the aspartic acid-arginine-tyrosine motif of the M2 muscarinic receptor, but not intracellular loop 3, significantly impaired the voltage-dependence of RGS4 function. Thus, our results demonstrated that voltage-dependence of RGS4 modulation is derived from the M2 muscarinic receptor. These results provide novel insights into how membrane potential impacts G-protein signaling by modulating GPCR communication with downstream effectors

RGS4 regulates partial agonism of the M2 muscarinic receptor-activated K+ currents.

Partial agonists are used clinically to avoid overstimulation of receptor-mediated signalling, as they produce a submaximal response even at 100% receptor occupancy. The submaximal efficacy of partial agonists is due to conformational change of the agonist-receptor complex, which reduces effector activation. In addition to signalling activators, several regulators help control intracellular signal transductions. However, it remains unclear whether these signalling regulators contribute to partial agonism. Here we show that regulator of G-protein signalling (RGS) 4 is a determinant for partial agonism of the M2 muscarinic receptor (M2R). In rat atrial myocytes, pilocarpine evoked smaller G-protein-gated K(+) inwardly rectifying (KG) currents than those evoked by ACh. In a Xenopus oocyte expression system, pilocarpine acted as a partial agonist in the presence of RGS4 as it did in atrial myocytes, while it acted like a full agonist in the absence of RGS4. Functional couplings within the agonist-receptor complex/G-protein/RGS4 system controlled the efficacy of pilocarpine relative to ACh. The pilocarpine-M2R complex suppressed G-protein-mediated activation of KG currents via RGS4. Our results demonstrate that partial agonism of M2R is regulated by the RGS4-mediated inhibition of G-protein signalling. This finding helps us to understand the molecular components and mechanism underlying the partial agonism of M2R-mediated physiological responses

Facilitation of I Kr current by some hERG channel blockers suppresses early afterdepolarizations.

Drug-induced block of the cardiac rapid delayed rectifying potassium current (I Kr), carried by the human ether-a-go-go-related gene (hERG) channel, is the most common cause of acquired long QT syndrome. Indeed, some, but not all, drugs that block hERG channels cause fatal cardiac arrhythmias. However, there is no clear method to distinguish between drugs that cause deadly arrhythmias and those that are clinically safe. Here we propose a mechanism that could explain why certain clinically used hERG blockers are less proarrhythmic than others. We demonstrate that several drugs that block hERG channels, but have favorable cardiac safety profiles, also evoke another effect; they facilitate the hERG current amplitude in response to low-voltage depolarization. To investigate how hERG facilitation impacts cardiac safety, we develop computational models of I Kr block with and without this facilitation. We constrain the models using data from voltage clamp recordings of hERG block and facilitation by nifekalant, a safe class III antiarrhythmic agent. Human ventricular action potential simulations demonstrate the ability of nifekalant to suppress ectopic excitations, with or without facilitation. Without facilitation, excessive I Kr block evokes early afterdepolarizations, which cause lethal arrhythmias. When facilitation is introduced, early afterdepolarizations are prevented at the same degree of block. Facilitation appears to prevent early afterdepolarizations by increasing I Kr during the repolarization phase of action potentials. We empirically test this prediction in isolated rabbit ventricular myocytes and find that action potential prolongation with nifekalant is less likely to induce early afterdepolarization than action potential prolongation with dofetilide, a hERG channel blocker that does not induce facilitation. Our data suggest that hERG channel blockers that induce facilitation increase the repolarization reserve of cardiac myocytes, rendering them less likely to trigger lethal ventricular arrhythmias

Reduction techniques of the back gate effect in the SOI Pixel Detector

We have fabricated monolithic pixel sensors in 0.2 μm Silicon-On-Insulator (SOI) CMOS technology, consisting of a thick sensor layer and a thin circuit layer with an insulating buried-oxide, which has many advantages. However, it has been found that the applied electric field in the sensor layer also affects the transistor operation in the adjacent circuit layer. This limits the applicable sensor bias well below the full depletion voltage. To overcome this, we performed a TCAD simulation and added an additional p-well (buried pwell) in the SOI process. Designs and preliminary results are presented

A new scheme for the running coupling constant in gauge theories using Wilson loops

We propose a new renormalization scheme of the running coupling constant in general gauge theories using the Wilson loops. The renormalized coupling constant is obtained from the Creutz ratio in lattice simulations and the corresponding perturbative coefficient at the leading order. The latter can be calculated by adopting the zeta-function resummation techniques. We perform a benchmark test of our scheme in quenched QCD with the plaquette gauge action. The running of the coupling constant is determined by applying the step-scaling procedure. Using several methods to improve the statistical accuracy, we show that the running coupling constant can be determined in a wide range of energy scales with relatively small number of gauge configurations.Comment: 30pages, figs and comments added,reference added(v3

Study of the conformal hyperscaling relation through the Schwinger-Dyson equation

We study corrections to the conformal hyperscaling relation in the conformal window of the large Nf QCD by using the ladder Schwinger-Dyson (SD) equation as a concrete dynamical model. From the analytical expression of the solution of the ladder SD equation, we identify the form of the leading mass correction to the hyperscaling relation. We find that the anomalous dimension, when identified through the hyperscaling relation neglecting these corrections, yields a value substantially lower than the one at the fixed point \gamma_m^* for large mass region. We further study finite-volume effects on the hyperscaling relation, based on the ladder SD equation in a finite space-time with the periodic boundary condition. We find that the finite-volume corrections on the hyperscaling relation are negligible compared with the mass correction. The anomalous dimension, when identified through the finite-size hyperscaling relation neglecting the mass corrections as is often done in the lattice analyses, yields almost the same value as that in the case of the infinite space-time neglecting the mass correction, i.e., a substantially lower value than \gamma_m^* for large mass. We also apply the finite-volume SD equation to the chiral-symmetry-breaking phase and find that when the theory is close to the critical point such that the dynamically generated mass is much smaller than the explicit breaking mass, the finite-size hyperscaling relation is still operative. We also suggest a concrete form of the modification of the finite-size hyperscaling relation by including the mass correction, which may be useful to analyze the lattice data.Comment: 17 pages, 9 figure

Meson masses in large Nf QCD from the Bethe-Salpeter equation

We solve the homogeneous Bethe-Salpeter (HBS) equation for the scalar, pseudoscalar, vector, and axial-vector bound states of quark and anti-quark in large Nf QCD with the improved ladder approximation in the Landau gauge. The quark mass function in the HBS equation is obtained from the Schwinger-Dyson (SD) equation in the same approximation for consistency with the chiral symmetry. Amazingly, due to the fact that the two-loop running coupling of large Nf QCD is explicitly written in terms of an analytic function, large Nf QCD turns out to be the first example in which the SD equation can be solved in the complex plane and hence the HBS equation directly in the time-like region. We find that approaching the chiral phase transition point from the broken phase, the scalar, vector, and axial-vector meson masses vanish to zero with the same scaling behavior, all degenerate with the massless pseudoscalar meson. This may suggest a new type of manifestation of the chiral symmetry restoration in large Nf QCD.Comment: 33 pages, 16 figures. Typos are corrected. Minor corrections and references are added. Version to appear in Phys. Rev.

Distinguishing among Technicolor/Warped Scenarios in Dileptons

Models of dynamical electroweak symmetry breaking usually include new spin-1 resonances, whose couplings and masses have to satisfy electroweak precision tests. We propose to use dilepton searches to probe the underlying structure responsible for satisfying these. Using the invariant mass spectrum and charge asymmetry, we can determine the number, parity, and isospin of these resonances. We pick three models of strong/warped symmetry breaking, and show that each model produces specific features that reflect this underlying structure of electroweak symmetry breaking and cancellations.Comment: Added missing referenc