Search

A system of dispersive representations of the Omnès–Khuri–Treiman–Sawyer–Wali type for the final-state interactions in the amplitudes of the K → π π π weak transitions is constructed, under the assumptions that CP and isospin symmetries are conserved. Both the Δ I = 1 / 2 and Δ I = 3 / 2 transition channels are considered. The set of single-variable functions involved in these representations is identified, and the polynomial ambiguities in their definitions are discussed. Numerical solutions for the system of coupled integral relations satisfied by these functions are provided, and the determination of the subtraction constants in terms of the experimental information on the amplitudes in the decay region is addressed. [ABSTRACT FROM AUTHOR]

We consider a left-right symmetric extension of the Standard Model where the spontaneous breakdown of the left-right symmetry is triggered by doublets. The electroweak ρ parameter is protected from large corrections in this Doublet Left-Right Model (DLRM), contrary to the triplet case. This allows in principle for more diverse patterns of symmetry breaking. We consider several constraints on the gauge and scalar sectors of DLRM: the unitarity of scattering processes involving gauge bosons with longitudinal polarisations, the radiative corrections to the muon ∆r parameter and the electroweak precision observables measured at the Z pole and at low energies. Combining these constraints within the frequentist CKMfitter approach, we see that the fit pushes the scale of left-right symmetry breaking up to a few TeV, while favouring an electroweak symmetry breaking triggered not only by the SU(2)L×SU(2)R bi-doublet, which is the case most commonly considered in the literature, but also by the SU(2)L doublet. [ABSTRACT FROM AUTHOR]

Several subtypes of modulatory neurons co‐express vesicular glutamate transporters (VGLUTs) in addition to their cognate vesicular transporters. These neurons are believed to establish new forms of neuronal communication. The atypical VGLUT3 is of particular interest since in the striatum this subtype is found in tonically active cholinergic interneurons (TANs) and in a subset of 5‐HT fibers. The striatum plays a major role in psychomotor effects induced by amphetamine. Whether and how VGLUT3‐operated glutamate/ACh or glutamate/5HT co‐transmissions modulates psychostimulants‐induced maladaptive behaviors is still unknown. Here, we investigate the involvement of VGLUT3 and glutamate co‐transmission in amphetamine‐induced psychomotor effects and stereotypies. Taking advantage of constitutive and cell‐type specific VGLUT3‐deficient mouse lines, we tackled the hypothesis that VGLUT3 could gate psychomotor effects (locomotor activity and stereotypies) induced by acute or chronic administration of amphetamine. Interestingly, VGLUT3‐null mice demonstrated blunted amphetamine‐induced stereotypies as well as reduced striatal ∆FosB expression. VGLUT3‐positive varicosities within the striatum arise in part from 5HT neurons. We tested the involvement of VGLUT3 deletion in serotoninergic neurons in amphetamine‐induced stereotypies. Mice lacking VGLUT3 specifically in 5HT fibers showed no alteration to amphetamine sensitivity. In contrast, specific deletion of VGLUT3 in cholinergic neurons partially phenocopied the effects observed in the constitutive knock‐out mice. Our results show that constitutive deletion of VGLUT3 modulates acute and chronic locomotor effects induced by amphetamine. They point to the fact that the expression of VGLUT3 in multiple brain areas is pivotal in gating amphetamine‐induced psychomotor adaptations. Open Science Badges: This article has received a badge for *Open Materials* because it provided all relevant information to reproduce the study in the manuscript. The complete Open Science Disclosure form for this article can be found at the end of the article. More information about the Open Practices badges can be found at https://cos.io/our-services/open-science-badges/. VGLUT3 is detected in neurons that use other neurotransmitters than glutamate, a neurochemical feature known as vesicular synergy. Here, we investigate the involvement of VGLUT3 in amphetamine‐induced psychomotor effects. We found that deletion of VGLUT3 promoted hyperlocomotor responses whereas blunted stereotypies were observed following amphetamine administration. A reduced expression of ∆FosB in the striatum mirrored such phenotype. In the striatum, VGLUT3 is expressed in cholinergic neurons and 5HT fibers. Cell‐type specific deletion of VGLUT3 in ACh and 5HT neurons revealed a complex dynamic regulation of amphetamine‐related effects. Our study points VGLUT3 as a critical modulator of psychostimulants‐induced adaptations. Open Science: This manuscript was awarded with the Open Materials Badge For more information see: https://cos.io/our-services/open-science-badges/ [ABSTRACT FROM AUTHOR]

The extraction of the B → K ⁎ transition form factors from lattice data is studied, applying non-relativistic effective field theory in a finite volume. The possible mixing of πK and ηK states is taken into account. The two-channel analogue of the Lellouch–Lüscher formula is reproduced. Due to the resonance nature of the K ⁎ , an equation is derived, which allows to determine the form factors at the pole position in a process-independent manner. The infinitely-narrow width approximation of the results is discussed. [ABSTRACT FROM AUTHOR]

Two aspects of isospin breaking in the decay $$K^\pm \rightarrow \pi ^0 \pi ^0 e^\pm \mathop {\nu _e}\limits ^{_{(-)}}$$ are studied and discussed. The first addresses the possible influence of the phenomenological description of the unitarity cusp on the extraction of the normalization of the form factor from data. Using the scalar form factor of the pion as a theoretical laboratory, we find that this determination is robust under variations of the phenomenological parameterizations of the form factor. The second aspect concerns the issue of radiative corrections. We compute the radiative corrections to the total decay rate for $$K^\pm \rightarrow \pi ^0 \pi ^0 e^\pm \mathop {\nu _e}\limits ^{_{(-)}}$$ in a setting that allows comparison with the way radiative corrections were handled in the channel $$K^\pm \rightarrow \pi ^+ \pi ^- e^\pm \mathop {\nu _e}\limits ^{_{(-)}}$$ . We find that, once radiative corrections are included, the normalizations of the form factor as determined experimentally from data in the two decay channels come to a better agreement. The remaining discrepancy can easily be accounted for by other isospin-breaking corrections, mainly those due to the difference between the masses of the up and down quarks. [ABSTRACT FROM AUTHOR]

Using the non-relativistic effective field theory framework in a finite volume, we discuss the extraction of the ΔNγ* transition form factors from lattice data. A counterpart of the Lüscher approach for the matrix elements of unstable states is formulated. In particular, we thoroughly discuss various kinematic settings, which are used in the calculation of the above matrix element on the lattice. The emerging Lüscher-Lellouch factor and the analytic continuation of the matrix elements into the complex plane are also considered in detail. A full group-theoretical analysis of the problem is made, including the partial-wave mixing and projecting out the invariant form factors from data. [ABSTRACT FROM AUTHOR]

Isospin breaking in the K form factors induced by the difference between charged and neutral pion masses is studied. Starting from suitably subtracted dispersion representations, the form factors are constructed in an iterative way up to two loops in the low-energy expansion by implementing analyticity, crossing, and unitarity due to two-meson intermediate states. Analytical expressions for the phases of the two-loop form factors of the K→ π π e ν channel are given, allowing one to connect the difference of form-factor phase shifts measured experimentally (out of the isospin limit) and the difference of S- and P-wave ππ phase shifts studied theoretically (in the isospin limit). The isospin-breaking correction consists of the sum of a universal part, involving only ππ rescattering, and a process-dependent contribution, involving the form factors in the coupled channels. The dependence on the two S-wave scattering lengths $a_{0}^{0}$ and $a_{0}^{2}$ in the isospin limit is worked out in a general way, in contrast to previous analyses based on one-loop chiral perturbation theory. The latter is used only to assess the subtraction constants involved in the dispersive approach. The two-loop universal and process-dependent contributions are estimated and cancel partially to yield an isospin-breaking correction close to the one-loop case. The recent results on the phases of K→ π π e ν form factors obtained by the NA48/2 collaboration at the CERN SPS are reanalysed including this isospin-breaking correction to extract values for the scattering lengths $a_{0}^{0}$ and $a_{0}^{2}$, as well as for low-energy constants and order parameters of two-flavour χPT. [ABSTRACT FROM AUTHOR]

Abstract: Acetylcholinesterase (AChE) terminates the action of acetylcholine at cholinergic synapses thereby preventing rebinding of acetylcholine to nicotinic postsynaptic receptors at the neuromuscular junction. Here we show that AChE is not localized close to these receptors on the postsynaptic surface, but is instead clustered along the presynaptic membrane and deep in the postsynaptic folds. Because AChE is anchored by ColQ in the basal lamina and is linked to the plasma membrane by a transmembrane subunit (PRiMA), we used a genetic approach to evaluate the respective contribution of each anchoring oligomer. By visualization and quantification of AChE in mouse strains devoid of ColQ, PRiMA or AChE, specifically in the muscle, we found that along the nerve terminus the vast majority of AChE is anchored by ColQ that is only produced by the muscle, whereas very minor amounts of AChE are anchored by PRiMA that is produced by motoneurons. In its synaptic location, AChE is therefore positioned to scavenge ACh that effluxes from the nerve by non-quantal release. AChE–PRiMA, produced by the muscle, is diffusely distributed along the muscle in extrajunctional regions. [ABSTRACT FROM AUTHOR]

Background: We wished to develop alternate production strategies to generate antibodies against traditionally problematic antigens. As a model we chose butyrylcholinesterase (BChE), involved in termination of cholinergic signaling, and widely considered as a poor immunogen. Methodology/Principal Findings: Jettisoning traditional laborious in silico searching methods to define putative epitopes, we simply immunized available BChE knock-out mice with full-length recombinant BChE protein (having been produced for crystallographic analysis). Immunization with BChE, in practically any form (recombinant human or mouse BChE, BChE purified from human serum, native or denatured), resulted in strong immune responses. Native BChE produced antibodies that favored ELISA and immunostaining detection. Denatured and reduced BChE were more selective for antibodies specific in Western blots. Two especially sensitive monoclonal antibodies were found capable of detecting 0.25 ng of BChE within one min by ELISA. One is specific for human BChE; the other cross-reacts with mouse and rat BChE. Immunization of wild-type mice served as negative controls. Conclusions/Significance: We examined a simple, fast, and highly efficient strategy to produce antibodies by mining two expanding databases: namely those of knock-out mice and 3D crystallographic protein-structure analysis. We conclude that the immunization of knock-out mice should be a strategy of choice for antibody production. [ABSTRACT FROM AUTHOR]

Abstract: A mouse strain with a deleted acetylcholinesterase (AChE) gene (AChE knockout) shows a decreased inspiration time and increased tidal volume and ventilation .To investigate the respective roles of AChE in brain and muscle, we recorded respiration by means of whole-body plethysmography in knockout mice with tissue selective deletions in AChE expression. A mouse strain with the anchoring domains of AChE deleted (del E5+6 knockout mice) has very low activity in the brain and neuromuscular junction, but increased monomeric AChE in serum. A mouse strain with deletion of the muscle specific region of AChE (del i1RR knockout mice) exhibits no expression in muscle, but unaltered expression in the central nervous system. Neither strain exhibits the pronounced phenotypic traits observed in the complete AChE knockout strain. A third strain lacking the anchor molecule PRiMA, has no functional AChE and butyrylcholinesterase (BChE) in brain and an unaltered respiratory function. BChE inhibition by bambuterol decreases tidal volume and body temperature in del E5+6 and i1RR knockout strains, but not in PRiMA deletion or wild-type controls. We find that: (1) deletion of the full AChE gene is required for a pronounced alteration in respiratory phenotype, (2) BChE is involved in respiratory muscles contraction and temperature control in del E5+6 and i1RR knockout mice, and (3) AChE expression requiring a gene product splice to either exons 5 and 6 or regulated by intron1 influences temperature control. [Copyright &y& Elsevier]

Abstract: We perform a matching of the two loop-chiral perturbation theory representation of the scalar Kπ form factor to a dispersive one. Knowing the value of and in the Standard Model (SM) allows to determine two LECs, the slope of the scalar form factor and the deviation of the Callan–Treiman theorem. Going beyond the SM and assuming the knowledge of the slope of the scalar form factor from experiment, the matching allows us to determine the ratio of , , a certain combination of non-standard couplings, the deviation of the Callan–Treiman theorem and the two LECs. [Copyright &y& Elsevier]

Abstract: We study the extraction of Δ-resonance parameters from lattice data for small quark masses, corresponding to the case of an unstable Δ. To this end, we calculate the spectrum of the correlator of two Δ-fields in a finite Euclidean box up-to-and-including in the small scale expansion using infrared regularization. On the basis of our numerical study, we argue that the extraction of the parameters of the Δ-resonance (in particular, of the mass and the pion–nucleon–delta coupling constant) from the measured volume dependence of the lowest energy levels should be feasible. [Copyright &y& Elsevier]

Abstract: Theoretical as well as experimental progress has been made in the last decade in describing the properties of baryons. In this review I will mostly report on the theoretical issues. Two non-perturbative methods are privileged frameworks for studying their properties in the low-energy domain: chiral perturbation theory, the effective field theory of the Standard Model at energies below 1 GeV and lattice QCD. I will mainly concentrate here on the first one but I will also discuss the complementarity of the two methods. Chiral extrapolations for lattice simulations of some nucleon properties will be investigated. I will then concentrate on processes involving at most two nucleons, describing for example pion–nucleon and pion–deuteron scattering, pion photo- and electro-production off the nucleon and the deuteron and doubly virtual Compton scattering. Three flavor calculations will also be reviewed. [Copyright &y& Elsevier]

This review gives a brief introduction to chiral perturbation theory in its various settings. We discuss some applications of recent interest, including chiral extrapolations for lattice gauge theory. [ABSTRACT FROM AUTHOR]

Abstract: At the neuromuscular junction (NMJ) acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) can hydrolyze acetylcholine (ACh). Released ACh quanta are known to diffuse rapidly across the narrow synaptic cleft and pairs of ACh molecules cooperate to open endplate channels. During their diffusion through the cleft, or after being released from muscle nicotinic ACh receptors (nAChRs), most ACh molecules are hydrolyzed by AChE highly concentrated at the NMJ. Advances in mouse genomics offered new approaches to assess the role of specific cholinesterases involved in synaptic transmission. AChE knockout mice (AChE-KO) provide a valuable tool for examining the complete abolition of AChE activity and the role of BChE. AChE-KO mice live to adulthood, and exhibit an increased sensitivity to BChE inhibitors, suggesting that BChE activity facilitated their survival and compensated for AChE function. Our results show that BChE is present at the endplate region of wild-type and AChE-KO mature muscles. The decay time constant of focally recorded miniature endplate currents was 1.04±0.06 ms in wild-type junctions and 5.4 ms±0.3 ms in AChE-KO junctions, and remained unaffected by BChE-specific inhibitors, indicating that BChE is not limiting ACh duration on endplate nAChRs. Inhibition of BChE decreased evoked quantal ACh release in AChE-KO NMJs. This reduction in ACh release can explain the greatest sensitivity of AChE-KO mice to BChE inhibitors. BChE is known to be localized in perisynaptic Schwann cells, and our results strongly suggest that BChE''s role at the NMJ is to protect nerve terminals from an excess of ACh. [Copyright &y& Elsevier]

Abstract: We analyze the nucleon axial-vector coupling to two loops in chiral perturbation theory. We show that chiral extrapolations based on this representation require lattice data with pion masses below 300 MeV. [Copyright &y& Elsevier]

Abstract: Clean tests of a small admixture of right-handed quark currents directly coupled to the standard W are still lacking. We show that such non-standard couplings can be significantly constrained measuring the value of the scalar Kπ form factor at the Callan–Treiman point to a few percent. A realistic prospect of such a measurement in decay based on an accurate dispersive representation of the scalar form factor is presented. The inadequacy of the currently used linear parametrization is explained and illustrated using recent KTeV data. We briefly comment on the charged kaon mode. [Copyright &y& Elsevier]

In vitro studies have widely demonstrated that the abundance and availability of G-protein-coupled receptors (GPCRs) at the cell surface is regulated by the neuronal environment and is the result of complex intraneuronal trafficking. However, this regulation is still poorly understood in vivo. Modulation of receptor availability at the neuronal membrane is a key event in the regulation of neuronal functions (e.g. neurotransmitter release or neuronal excitability in physiological, pathological or therapeutic conditions). We discuss the effects of duration of receptor stimulation (acute versus chronic) on the intraneuronal trafficking of GPCRs in vivo, and we show that this trafficking might differ according to subcellular compartment (soma, dendrites or axon terminals). [Copyright &y& Elsevier]

Abstract: We calculate the nucleon and the delta mass to fourth order in a covariant formulation of the small scale expansion. We analyze lattice data from the MILC Collaboration and demonstrate that the available lattice data combined with our knowledge of the physical values for the nucleon and delta masses lead to consistent chiral extrapolation functions for both observables up to fairly large pion masses. This holds in particular for very recent data on the delta mass from the QCDSF Collaboration. The resulting pion–nucleon sigma term is . This first quantitative analysis of the quark-mass dependence of the structure of the in full QCD within chiral effective field theory suggests that (the real part of) the nucleon–delta mass-splitting in the chiral limit, , is slightly larger than at the physical point. Further analysis of simultaneous fits to nucleon and delta lattice data are needed for a precision determination of the properties of the first excited state of the nucleon. [Copyright &y& Elsevier]

Abstract: The shape of the electron energy spectrum in 3H β-decay permits a direct assay of the absolute scale of the neutrino mass; a highly accurate theoretical description of the electron energy spectrum is necessary to the empirical task. We update Sirlin''s calculation of the outer radiative correction to nuclear β-decay to take into account the non-zero energy resolution of the electron detector. In previous 3H β-decay studies the outer radiative corrections were neglected all together; only Coulomb corrections to the spectrum were included. This neglect artificially pushes in a potentially significant way. We present a computation of the theoretical spectrum appropriate to the extraction of the neutrino mass in the sub-eV regime. [Copyright &y& Elsevier]

We consider radiative β-decay of the neutron in heavy baryon chiral perturbation theory, with an extension including explicit Δ degrees of freedom. We compute the photon energy spectrum as well as the photon polarization; both observables are dominated by the electron bremsstrahlung contribution. Nucleon-structure effects not encoded in the weak coupling constants gA and gV are determined at next-to-leading order in the chiral expansion, and enter at the O(0.5%)-level, making a sensitive test of the Dirac structure of the weak currents possible. [Copyright &y& Elsevier]

We discuss the use of cutoff methods in chiral perturbation theory. We develop a cutoff scheme based on the operator structure of the effective field theory that allows to suppress high momentum contributions in Goldstone boson loop integrals and by construction is free of the problems traditional cutoff schemes have with gauge invariance or chiral symmetries. As an example, we discuss the chiral expansion of the nucleon mass. We show that the mass of a nucleon in heavy baryon chiral perturbation theory has a well behaved chiral expansion up to effective Goldstone boson masses of 400 MeV when one utilizes standard dimensional regularization techniques. With the help of the here developed cutoff scheme we can demonstrate a well-behaved chiral expansion for the nucleon mass up to 600 MeV of effective Goldstone boson masses. We also discuss in detail the price, in numbers of additional short distance operators involved, that has to be paid for this extended range of applicability of chiral perturbation theory with cutoff regularization. We also compare the fourth-order result for the chiral expansion of the nucleon mass with lattice results and draw some conclusions about chiral extrapolations based on such type of representation. [Copyright &y& Elsevier]

We have studied ”in vivo” neurochemically identified striatal neurons to analyze the localisation and the trafficking of dopamine and acetylcholine G protein coupled receptors (GPCR) (D1R, D2R, m2R and m4R) under the influence of neurotransmitter environment. We have identified receptors in tissue sections through immunohistochemical detection at the light and electron microscopic level. We have identified receptors in normal animals and after acute and chronic stimulations. We have quantified receptors through image analysis at the electron microscopic level in relation to various subcellular compartments. Our results demonstrate that, in normal conditions, GPCRs are mostly associated with plasma membrane of the striatal neurons, mostly at extra-synaptic sites. In certain instances (m4R; D2R), receptors have prominent localisation inside the rough endoplasmic reticulum. Our results also show that two distinct receptors for a same neurotransmitter may have distinct subcellular localisation in a same neuronal population (m2R versus m4R) and that the same neurotransmitter receptor (m4R) can have distinct localisation in distinct neuronal populations (cytoplasm versus cell surface). After acute stimulation, cell surface receptors undergo dramatic subcellular changes that involve plasma membrane depletion, internalisation in endosomes and in multivesicular bodies. Such changes are reversible after the end of the stimulation and are blocked by antagonist action. Chronic stimulation also provokes changes in subcellular localisation with specific pattern: plasma membrane depletion, and exaggerated storage of receptors in rough endoplasmic reticulum and eventually Golgi complex (D1R; m2R and m4R). Decreasing chronic receptor stimulation reverses such changes. These results demonstrate that, ”in vivo”, in the striatum, GPCRs undergo complex intraneuronal trafficking under the influence of neurochemical environment in conditions that dramatically modulate the number of cell surface receptors available for interaction with neurotransmitters or drugs. This confirms that ”in vivo”, the trafficking and the subcellular compartmentalization of GPCRs may contribute to regulate neuronal sensitivity and neuronal interactions in physiological, experimental and pathological conditions, including in therapeutic conditions. [Copyright &y& Elsevier]

We present a Lorentz-invariant formulation of baryon chiral perturbation theory including spin-3/2 fields. Particular attention is paid to the projection on the spin-3/2 components of the delta fields. We also discuss the nucleon mass and the pion–nucleon sigma term. [Copyright &y& Elsevier]

We have studied the consequences of the constitutive acetylcholinesterase (AChE) deficiency in knockout mice for the AChE gene on the subcellular localization of the m2 receptor (m2R) and the regulation of its intraneuronal compartmentalization by the cholinergic environment, using immunohistochemistry at light and electron microscopic levels. (1) In AChE +/+ mice in vivo, m2R is mainly located at the neuronal membrane in striatum, hippocampus, and cortex. In AChE −/− mice, m2R is almost absent at the membrane but is accumulated in the endoplasmic reticulum and Golgi complex. (2) In vivo and in vitro (organotypic culture) dynamic studies demonstrate that the balance between membrane and intracytoplasmic m2R can be regulated by the cholinergic influence: In AChE −/− mice, m2R is translocated from the cytoplasm to the cell surface after (1) blockade of muscarinic receptors by atropine, (2) supplementation of AChE −/− neurons with AChE in vitro, and (3) disruption of the cortical and hippocampal cholinergic afferents in vitro. Our results suggest that the neurochemical environment may contribute to the control of the abundance and availability of cell surface receptors, and consequently to the control of neuronal sensitivity to neurotransmitters or drugs, by regulating their delivery from the endoplasmic reticulum and Golgi complex. [Copyright &y& Elsevier]

We study the chiral loop corrections to the generalized Gerasimov–Drell–Hearn sum rule of the nucleon for finite photon virtuality in the framework of a Lorentz-invariant formulation of baryon chiral perturbation theory. We perform a complete one-loop calculation and obtain significant differences to previously found results based on the heavy baryon approach for the proton and neutron spin-dependent forward Compton amplitudes. [Copyright &y& Elsevier]

Acetylcholinesterase inhibitors (AChE-I) of various pharmacological classes have been used to provoke acute and chronic hypercholinergy in brain. Each condition induces a dramatic decrease of the abundance of muscarinic receptors at the membrane of neurons with simultaneous increase of these receptors in the cytoplasm in association with different subcellular organelles with characteristics depending on the duration of the treatment (short-term versus long term treatment). Each condition also induces a dramatic increase of cytoplasmic receptors associated with endosomes and multivesicular bodies. Chronic treatment with MTF induces a general decrease of m4R in the striatum without modification of the mRNA level but with an exaggerated abundance of muscarinic receptors in the cytoplasm at the sites of synthesis and maturation, i.e., endoplasmic reticulum, nuclear membrane and Golgi apparatus. These results suggest that the membrane abundance and intraneuronal distribution of neurotransmitter receptors are modified following drug treatment with specificity depending on the nature and the duration of treatment. [Copyright &y& Elsevier]

AbstractGlutamatergic neurotransmission in the neostriatum and the globus pallidus is mediated through NMDA-type as well as other glutamate receptors and is critical in the expression of basal ganglia function. In order to characterize the cellular, subcellular and subsynaptic localization of NMDA receptors in the neostriatum and globus pallidus, multiple immunocytochemical techniques were applied using antibodies that recognize the NR1 subunit of the NMDA receptor. In order to determine the spatial relationship between NMDA receptors and AMPA receptors, double labelling was performed with the NR1 antibodies and an antibody that recognizes the GluR2 and 3 subunits of the AMPA receptor. In the neostriatum all neurons with characteristics of spiny projection neurons, some interneurons and many dendrites and spines were immunoreactive for NR1. In the globus pallidus most perikarya and many dendritic processes were immunopositive. Immunogold methods revealed that most NR1 labelling is associated with asymmetrical synapses and, like the labelling for GluR2/3, is evenly spread across the synapse. Double immunolabelling revealed that in neostriatum, over 80% of NR1-positive axospinous synapses are also positive for GluR2/3. In the globus pallidus most NR1-positive synapses are positive for GluR2/3. In both regions many synapses labelled only for GluR2/3 were also detected. These results, together with previous data, suggest that NMDA and AMPA receptor subunits are expressed by the same neurons in the neostriatum and globus pallidus and that NMDA and AMPA receptors are, at least in part, colocalized at individual asymmetrical synapses. The synaptic responses to glutamate in these regions are thus likely be mediated by both AMPA and NMDA receptors at the level of individual synapses. [ABSTRACT FROM AUTHOR]

Maitotoxin, a potent marine toxin extracted from peredinians, was found to mimic fertilization in Xenopus oocytes and to trigger the breakdown of phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2, the precursor of inositol 1,4,5-trisphosphate], an increase of intracellular pCa and the cortical reaction, including the exocytosis of cortical granules and a wave-like propagation of contraction in the animal hemisphere. All these effects of maitotoxin required the presence of external calcium. Moreover, the toxin considerably increased 24 influx in amphibian oocytes arrested at first meiotic prophase, due to the permanent activation of voltage-dependent Ca2+ channels. Nevertheless it is doubtful that maitotoxin acts primarily as a Ca2+ ionophore or at the level of Ca2+ channels. Indeed no stimulation of Ca2+ uptake was observed in metaphase-II-arrested oocytes, although maitotoxin readily triggered the breakdown of PtdIns(4,5)P2 as well as the cortical reaction in such cells. On the other hand, PtdIns(4,5)P2 breakdown was not reduced in oocytes rnicroinjected with EGTA, although the calcium chelator prevented the oocytes from undergoing the cortical reaction. Taken together, these findings support the view that the toxin might act primarily by increasing PtdIns(4,5)P2 phosphodiesterase activity. [ABSTRACT FROM AUTHOR]

Loss-of-function mutations in the potassium-chloride cotransporter KCC3 lead to Andermann syndrome, a severe sensorimotor neuropathy characterized by areflexia, amyotrophy and locomotor abnormalities. The molecular events responsible for axonal loss remain poorly understood. Here, we establish that global or neuron-specific KCC3 loss-of-function in mice leads to early neuromuscular junction (NMJ) abnormalities and muscular atrophy that are consistent with the pre-synaptic neurotransmission defects observed in patients. KCC3 depletion does not modify chloride handling, but promotes an abnormal electrical activity among primary motoneurons and mislocalization of Na + /K + -ATPase α1 in spinal cord motoneurons. Moreover, the activity-targeting drug carbamazepine restores Na + /K + -ATPase α1 localization and reduces NMJ denervation in Slc12a6 −/− mice. We here propose that abnormal motoneuron electrical activity contributes to the peripheral neuropathy observed in Andermann syndrome. [ABSTRACT FROM AUTHOR]

Two near infra-red (NIR) fluorescent probes HupNIR1 and HupNIR2 based on the huprine scaffold and cyanine 5.0 dye have been synthesised and evaluated in situ for the detection of acetylcholinesterases in different tissues. As anticipated by the initial properties of huprine, both probes displayed a high affinity and selectivity for AChE toward BChE, with IC50 values in the nanomolar range and without any non-specific binding in the tissues. HupNIR2 appears the best probe for AChE with a great selectivity and sensitivity for AChE even in the brain region displaying a low AChE concentration as striatum. Moreover, the binding of HupNIR2 is affected when AChE is inhibited with toxic molecules such as organophosphates. This work provides a new tool to visualize active AChE in biological applications. [ABSTRACT FROM AUTHOR]

In many brain areas, few cholinergic synapses are identified. Acetylcholine is released into the extracellular space and acts through diffuse transmission. Motoneurons, however, are contacted by numerous cholinergic terminals, indicating synaptic cholinergic transmission on them. The muscarinic m2 receptor is the major acetylcholine receptor subtype of motoneurons; therefore, we analyzed the localization of the m2 receptor in correlation with synapses by electron microscopic immunohistochemistry in the mouse trigeminal, facial, and hypoglossal motor nuclei. In all nuclei, m2 receptors were localized at the membrane of motoneuronal perikarya and dendrites. The m2 receptors were concentrated at cholinergic synapses located on the perikarya and most proximal dendrites. However, m2 receptors at cholinergic synapses represented only a minority (<10%) of surface m2 receptors. The m2 receptors were also enriched at glutamatergic synapses in both motoneuronal perikarya and dendrites. A relatively large proportion (20-30%) of plasma membrane-associated m2 receptors were located at glutamatergic synapses. In conclusion, the effect of acetylcholine on motoneuron populations might be mediated through a synaptic as well as diffuse type of transmission. J. Comp. Neurol. 521:2008-2024, 2013. © 2012 Wiley Periodicals, Inc. [ABSTRACT FROM AUTHOR]

CLIPR-59 is a new member of the cytoplasmic linker proteins (CLIP) family mainly localized to the trans-Golgi network. We show here that Clipr-59 expression in mice is restricted to specific pools of neurons, in particular motoneurons (MNs), and progressively increases from embryonic day 12.5 (E12.5) until the first postnatal days. We generated a Clipr-59 knockout mouse model that presents perinatal lethality due to respiratory defects. Physiological experiments revealed that this altered innervation prevents the normal nerveelicited contraction of the mutant diaphragm that is reduced both in amplitude and fatigue-resistance at E18.5, despite unaffected functional muscular contractility. Innervation of the mutant diaphragm is not altered until E15.5, but is then partially lost in the most distal parts of the muscle. Ultrastructural observations of neuromuscular junctions (NMJs) in the distal region of the diaphragm reveal a normal organization, but a lower density of nerve terminals capped by terminal Schwann cells in E18.5 mutant when compared with control embryos. Similar defects in NMJ stability, with a hierarchy of severity along the caudo-rostral axis, are also observed in other muscles innervated by facial and spinal MNs in Clipr-59 mutant mice. Clipr-59 deficiency therefore affects axon maintenance but not axon guidance toward muscle targets. Thus, CLIPR-59 is involved in the stabilization of specific motor axons at the NMJ during mouse late embryogenesis and its role is crucial for mouse perinatal development. [ABSTRACT FROM AUTHOR]

Increasing evidence suggests that somatostatin may control neuronal excitability during epileptogenesis. In the hippocampus, sst2A receptors are likely to mediate somatostatin inhibitory actions but little is known about their status in kindled tissues. In the present study, sst2A receptor and somatostatin immunoreactivity were examined by confocal microscopy in the hippocampus during and after kindling acquisition. In control rats, somatostatin-positive axon terminals were mainly found in the stratum lacunosum moleculare of CA1 area and in the outer molecular layer of the dentate gyrus. sst2A receptor immunoreactivity was diffusely distributed in the strata radiatum and oriens of CA1 and in the stratum moleculare of the dentate gyrus. Immunogold electron microscopy revealed that sst2A receptors were predominantly localized postsynaptically, at the plasma membrane of dendritic shafts and spines of principal neurons. During kindling epileptogenesis, qualitative and semiquantitative analysis revealed a progressive decrease of sst2A immunoreactivity in the outer molecular layer, which was spatially associated with an increase in somatostatin immunoreactivity. No obvious changes in sst2A receptor immunoreactivity were observed in other hippocampal subfields. These results suggest that the decrease of sst2A receptor immunoreactivity in the outer molecular layer reflects receptor down-regulation in distal dendrites of granule cells in response to chronic somatostatin release. Because the sst2A receptor appears to mediate anticonvulsant and antiepileptogenic effects of somatostatin, this may represent a pivotal mechanism contributing to epileptogenesis. [ABSTRACT FROM AUTHOR]

Striatal cholinergic interneurons ( CIN) are pivotal for the regulation of the striatal network. Acetylcholine ( ACh) released by CIN is centrally involved in reward behavior as well as locomotor or cognitive functions. Recently, BAC transgenic mice expressing channelrhodopsin-2 (ChR2) protein under the control of the choline acetyltransferase (Ch AT) promoter (Ch AT-ChR2) and displaying almost 50 extra copies of the VAChT gene were used to dissect cholinergic circuit connectivity and function using optogenetic approaches. These mice display over-expression of the vesicular acetylcholine transporter ( VAChT) and increased cholinergic tone. Consequently, Ch AT-ChR2 mice are a valuable model to investigate hypercholinergic phenotypes. Previous experiments established that Ch AT-ChR2 mice display an increased sensitivity to amphetamine induced-locomotor activity and stereotypes. In the present report, we analyzed the impact of VAChT over-expression in the striatum of Ch AT-ChR2 mice. Ch AT-ChR2 mice displayed increased locomotor sensitization in response to low dose of cocaine. In addition, we observed a dramatic remodeling of the morphology of CIN in Ch AT-ChR2 transgenic mice. VAChT immunolabeling was markedly enhanced in the soma and terminal of CIN from Ch AT-ChR2 mice as previously shown (Crittenden et al. 2014). Interestingly, the number of cholinergic varicosities was markedly reduced (−87%) whereas their size was significantly increased (+177%). Moreover, VAChT over-expression dramatically modified its trafficking along the somatodendritic and axonal arbor. These findings demonstrate that Ch AT-ChR2 mice present major alterations of CIN neuronal morphology and increased behavioral sensitization to cocaine, supporting the notion that the increased levels of VAChT observed in these mice make them fundamentally different from wild-type mice. [ABSTRACT FROM AUTHOR]

Within the hippocampus, the major somatostatin (SRIF) receptor subtype, the sst2A receptor, is localized at postsynaptic sites of the principal neurons where it modulates neuronal activity. Following agonist exposure, this receptor rapidly internalizes and recycles slowly through the trans-Golgi network. In epilepsy, a high and chronic release of somatostatin occurs, which provokes, in both rat and human tissue, a decrease in the density of this inhibitory receptor at the cell surface. The insulin-regulated aminopeptidase (IRAP) is involved in vesicular trafficking and shares common regional distribution with the sst2A receptor. In addition, IRAP ligands display anticonvulsive properties. We therefore sought to assess by in vitro and in vivo experiments in hippocampal rat tissue whether IRAP ligands could regulate the trafficking of the sst2A receptor and, consequently, modulate limbic seizures. Using pharmacological and cell biological approaches, we demonstrate that IRAP ligands accelerate the recycling of the sst2A receptor that has internalized in neurons in vitro or in vivo. Most importantly, because IRAP ligands increase the density of this inhibitory receptor at the plasma membrane, they also potentiate the neuropeptide SRIF inhibitory effects on seizure activity. Our results further demonstrate that IRAP is a therapeutic target for the treatment of limbic seizures and possibly for other neurological conditions in which downregulation of G-protein-coupled receptors occurs. [ABSTRACT FROM AUTHOR]

Terminal Schwann cells (TSCs) are key components of the mammalian neuromuscular junction (NMJ). How the TSCs sense the synaptic activity in physiological conditions remains unclear. We have taken advantage of the distinct localization of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) at the NMJ to bring out the function of different ACh receptors (AChRs). AChE is clustered by the collagen Q in the synaptic cleft and prevents the repetitive activation of muscle nicotinic AChRs. We found that BChE is anchored at the TSC by a proline-rich membrane anchor, the small transmembrane protein anchor of brain AChE. When BChE was specifically inhibited, ACh release was significant depressed through the activation of α7 nAChRs localized on the TSC and activated by the spillover of ACh. When both AChE and BChE were inhibited, the spillover increased and induced a dramatic reduction of ACh release that compromised the muscle twitch triggered by the nerve stimulation, α7 nAChRs at the TSC may act as a sensor for spillover of ACh adjusted by BChE and may represent an extrasynaptic sensor for homeostasis at the NMJ. In myasthenic rats, selective inhibition of AChE is more effective in rescuing muscle function than the simultaneous inhibition of AChE and BChE because the concomitant inhibition of BChE counteracts the positive action of AChE inhibition. These results show that inhibition of BChE should be avoided during the treatment of myasthenia and the pharmacological reversal of residual curarization after anesthesia. [ABSTRACT FROM AUTHOR]

For decades, neuroscientists have used enriched preparations of synaptic particles called synaptosomes to study synapse function. However, the interpretation of corresponding data is problematic as synaptosome preparations contain multiple types of synapses and non-synaptic neuronal and glial contaminants. We established a novel Fluorescence Activated Synaptosome Sorting ( FASS) method that substantially improves conventional synaptosome enrichment protocols and enables high-resolution biochemical analyses of specific synapse subpopulations. Employing knock-in mice with fluorescent glutamatergic synapses, we show that FASS isolates intact ultrapure synaptosomes composed of a resealed presynaptic terminal and a postsynaptic density as assessed by light and electron microscopy. FASS synaptosomes contain bona fide glutamatergic synapse proteins but are almost devoid of other synapse types and extrasynaptic or glial contaminants. We identified 163 enriched proteins in FASS samples, of which FXYD6 and Tpd52 were validated as new synaptic proteins. FASS purification thus enables high-resolution biochemical analyses of specific synapse subpopulations in health and disease. [ABSTRACT FROM AUTHOR]

Abstract: Schwartz-Jampel syndrome (SJS) is a recessive disorder with muscle hyperactivity that results from hypomorphic mutations in the perlecan gene, a basement membrane proteoglycan. Analyses done on a mouse model have suggested that SJS is a congenital form of distal peripheral nerve hyperexcitability resulting from synaptic acetylcholinesterase deficiency, nerve terminal instability with preterminal amyelination, and subtle peripheral nerve changes. We investigated one adult patient with SJS to study this statement in humans. Perlecan deficiency due to hypomorphic mutations was observed in the patient biological samples. Electroneuromyography showed normal nerve conduction, neuromuscular transmission, and compound nerve action potentials while multiple measures of peripheral nerve excitability along the nerve trunk did not detect changes. Needle electromyography detected complex repetitive discharges without any evidence for neuromuscular transmission failure. The study of muscle biopsies containing neuromuscular junctions showed well-formed post-synaptic element, synaptic acetylcholinesterase deficiency, denervation of synaptic gutters with reinnervation by terminal sprouting, and long nonmyelinated preterminal nerve segments. These data support the notion of peripheral nerve hyperexcitability in SJS, which would originate distally from synergistic actions of peripheral nerve and neuromuscular junction changes as a result of perlecan deficiency. [Copyright &y& Elsevier]

A correction to the article "CLIPR-59: a protein essential for neuromuscular junction stability during mouse late embryonic development" that was published in the previous issue is presented.

Abstract: The complete knockout of the acetylcholinesterase gene (AChE) in the mouse yielded a surprising phenotype that could not have been predicted from deletion of the cholinesterase genes in Drosophila, that of a living, but functionally compromised animal. The phenotype of this animal showed a sufficient compromise in motor function that precluded precise characterization of central and peripheral nervous functional deficits. Since AChE in mammals is encoded by a single gene with alternative splicing, additional understanding of gene expression might be garnered from selected deletions of the alternatively spliced exons. To this end, transgenic strains were generated that deleted exon 5, exon 6, and the combination of exons 5 and 6. Deletion of exon 6 reduces brain AChE by 93% and muscle AChE by 72%. Deletion of exon 5 eliminates AChE from red cells and the platelet surface. These strains, as well as knockout strains that selectively eliminate the AChE anchoring protein subunits PRiMA or ColQ (which bind to sequences specified by exon 6) enabled us to examine the role of the alternatively spliced exons responsible for the tissue disposition and function of the enzyme. In addition, a knockout mouse was made with a deletion in an upstream intron that had been identified in differentiating cultures of muscle cells to control AChE expression. We found that deletion of the intronic regulatory region in the mouse essentially eliminated AChE in muscle and surprisingly from the surface of platelets. The studies generated by these knockout mouse strains have yielded valuable insights into the function and localization of AChE in mammalian systems that cannot be approached in cell culture or in vitro. [ABSTRACT FROM AUTHOR]

We report the case of a congenital myasthenic syndrome due to a mutation in AGRN, the gene encoding agrin, an extracellular matrix molecule released by the nerve and critical for formation of the neuromuscular junction. Gene analysis identified a homozygous missense mutation, c.5125G>C, leading to the p.Gly1709Arg variant. The muscle-biopsy specimen showed a major disorganization of the neuromuscular junction, including changes in the nerve-terminal cytoskeleton and fragmentation of the synaptic gutters. Experiments performed in nonmuscle cells or in cultured C2C12 myotubes and using recombinant mini-agrin for the mutated and the wildtype forms showed that the mutated form did not impair the activation of MuSK or change the total number of induced acetylcholine receptor aggregates. A solid-phase assay using the dystrophin glycoprotein complex showed that the mutation did not affect the binding of agrin to α-dystroglycan. Injection of wild-type or mutated agrin into rat soleus muscle induced the formation of nonsynaptic acetylcholine receptor clusters, but the mutant protein specifically destabilized the endogenous neuromuscular junctions. Importantly, the changes observed in rat muscle injected with mutant agrin recapitulated the pre- and post-synaptic modifications observed in the patient. These results indicate that the mutation does not interfere with the ability of agrin to induce post-synaptic structures but that it dramatically perturbs the maintenance of the neuromuscular junction. [ABSTRACT FROM AUTHOR]

Acetylcholinesterase (AChE) accumulates on axonal varicosities and is primarily found as tetramers associated with a proline-rich membrane anchor (PRiMA). PRiMA is a small transmembrane protein that efficiently transforms secreted AChE to an enzyme anchored on the outer cell surface. Surprisingly, in the striatum of the PRiMA knock-out mouse, despite a normal level of AChEmRNA,we find only 2-3% of wild type AChE activity, with the residual AChE localized in the endoplasmic reticulum, demonstrating that PRiMA in vivo is necessary for intracellular processing of AChE in neurons. Moreover, deletion of the retention signal of the AChE catalytic subunit in mice, which is the domain of interaction with PRiMA, does not restore AChE activity in the striatum, establishing thatPRiMAis necessary to target and/or to stabilize nascent AChE in neurons. These unexpected findings open new avenues to modulating AChE activity and its distribution in CNS disorders. [ABSTRACT FROM AUTHOR]

Understanding the trafficking of G-protein-coupled receptors (GPCRs) is of particular importance, especially when modifications of the neurochemic environment occur as in pathological or therapeutic circumstances. In the central nervous system, although some GPCRs were reported to internalize in vivo, little is known about their trafficking downstream of the endocytic event. To address this issue, distribution and expression pattern of the major somatostatin receptor subtype, the somatostatin type 2 (sst2), was monitored in the hippocampus using immunofluorescence, autoradiographic and immunogold experiments from 10 minutes to 7 days after in vivo injection of the receptor agonist octreotide. We then analyzed whether postendocytic trafficking of the receptor was dependent upon integrity of the microtubule network using colchicine-injected animals. Together, our results suggest that upon agonist stimulation, dendritic receptors are retrogradely transported through a microtubule-dependent mechanism to a trans Golgi domain enriched in the t-SNARE syntaxin 6 and trans Golgi network 38 proteins, before recycling. Because we show that the exit rate from the trans Golgi apparatus back to the plasma membrane (hours) is slower than the entry rate (minutes), the neuronal postendocytic trafficking of sst2 receptor is likely to have functional consequences in several neurological diseases in which an increase in somatostatin release occurs. [ABSTRACT FROM AUTHOR]

Abstract: By using immunohistochemical approaches at the light and electron microscopic levels, we have shown that aging modifies the subcellular distribution of the m2 muscarinic autoreceptor (m2R) differentially at somato-dendritic postsynaptic sites and at axonal presynaptic sites in cholinergic basalocortical neurons, in vivo. In cholinergic perikarya and dendrites of the nucleus basalis magnocellularis (NBM), aging is associated with a decrease of the density of m2R at the plasma membrane and in the cytoplasm, suggesting a decrease of the total number of m2R in the somato-dendritic field. In contrast, the number of substance P receptors per somato-dendritic surface was not affected. In the frontal cortex (FC), we have shown a decrease of cytoplasmic m2R density also leading to a decrease of the number of m2R per surface of varicosities but with no change of the density of m2R at the membrane. Our results suggest that the decrease of m2R in the somato-dendritic field of the NBM, but not a modification of the number of presynaptic m2 autoreceptors at the plasma membrane in the FC, could contribute to the decrease of the efficacy of cholinergic transmission observed with aging in the rat. [Copyright &y& Elsevier]