Dopamine D 4 Receptor-Deficient Mice Display Cortical Hyperexcitability

The dopamine D(4) receptor (D(4)R) is predominantly expressed in the frontal cortex (FC), a brain region that receives dense input from midbrain dopamine (DA) neurons and is associated with cognitive and emotional processes. However, the physiological significance of this dopamine receptor subtype has been difficult to explore because of the slow development of D(4)R agonists and antagonists the selectivity and efficacy of which have been rigorously demonstrated in vivo. We have attempted to overcome this limitation by taking a multidimensional approach to the characterization of mice completely deficient in this receptor subtype. Electrophysiological current and voltage-clamp recordings were performed in cortical pyramidal neurons from wild-type and D(4)R-deficient mice. The frequency of spontaneous synaptic activity and the frequency and duration of paroxysmal discharges induced by epileptogenic agents were increased in mutant mice. Enhanced synaptic activity was also observed in brain slices of wild-type mice incubated in the presence of the selective D(4)R antagonist PNU-101387G. Consistent with greater electrophysiological activity, nerve terminal glutamate density associated with asymmetrical synaptic contacts within layer VI of the motor cortex was reduced in mutant neurons. Taken together, these results suggest that the D(4)R can function as an inhibitory modulator of glutamate activity in the FC.Fil: Rubinstein, Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Cepeda, Carlos. University of California at Los Angeles; Estados UnidosFil: Hurst, Raymond S.. University of California at Los Angeles; Estados UnidosFil: Flores Hernandez, Jorge. University of California at Los Angeles; Estados UnidosFil: Ariano, Marjorie A.. The Chicago Medical School; Estados UnidosFil: Falzone, Tomas Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Kozell, Laura B.. Oregon Health Sciences University; Estados UnidosFil: Meshul, Charles K.. Oregon Health Sciences University; Estados UnidosFil: Bunzow, James R.. Oregon Health Sciences University; Estados UnidosFil: Low, Malcolm J.. Oregon Health Sciences University; Estados UnidosFil: Levine, Michael S.. University of California at Los Angeles; Estados UnidosFil: Grandy, David K.. Oregon Health Sciences University; Estados Unido

Chemical imaging by dissolution analysis (CIDA): Localized kinetics of dissolution behavior to provide 2D chemical mapping and tomographic imaging on a nanoscale

A new approach to achieving chemical mapping on a nanoscale is described that can provide 2D and tomographic images of surface and near-surface structure. The method comprises dissolving material from the surface of the sample by applying a series of aliquots of solvent then analyzing their contents after removing them, in between exposures the surface is imaged with atomic force microscopy. This technique relies on being able to compensate for any drift between images by use of software. It was applied to a blend of two polymers, PMMA and PS. The analytical data identified the material that was dissolved and the topography images enabled the location of the various materials to be determined by analyzing local dis-solution kinetics. The prospects for generalizing the approach are discussed

Transcranial alternating current stimulation to the inferior parietal lobe decreases Mu suppression to egocentric, but not allocentric hand movements

Egocentric vs. allocentric perspective during observation of hand movements has been related to self-other differentiation such that movements observed from an egocentric viewpoint have been considered as self-related while movements observed from an allocentric viewpoint have been considered as belonging to someone else. Correlational studies have generally found that egocentric perspective induces greater neurophysiological responses and larger behavioural effects compared to an allocentric perspective. However, recent studies question previous findings by reporting greater (μ) suppression and greater transcranial magnetic stimulation (TMS) induced motor-evoked potentials (MEPs) during observation of allocentric compared to egocentric movements. Furthermore, self-other differentiation has been generally related to activity within the inferior parietal lobe (IPL), but direct evidence for a causal and functional role of IPL in self-other differentiation is lacking. The current study was therefore designed to investigate the influence that IPL exerts on self-other differentiation. To this aim, we measured the impact of individually adjusted alpha-tuned transcranial alternating current stimulation (tACS) applied over IPL on μ-suppression during hands movement observation from an egocentric and allocentric perspective. Electroencephalography (EEG) was recorded during movement observation before and immediately after tACS. Results demonstrated that tACS decreased μ-reactivity over sensorimotor (but not visual) regions for egocentric (but not allocentric) movement observation providing direct evidence for a causal involvement of IPL in the observation of self- but not other-related hands movement

Differential Modulation of Beta-Adrenergic Receptor Signaling by Trace Amine-Associated Receptor 1 Agonists

Trace amine-associated receptors (TAAR) are rhodopsin-like G-protein-coupled receptors (GPCR). TAAR are involved in modulation of neuronal, cardiac and vascular functions and they are potentially linked with neurological disorders like schizophrenia and Parkinson's disease. Subtype TAAR1, the best characterized TAAR so far, is promiscuous for a wide set of ligands and is activated by trace amines tyramine (TYR), phenylethylamine (PEA), octopamine (OA), but also by thyronamines, dopamine, and psycho-active drugs. Unfortunately, effects of trace amines on signaling of the two homologous β-adrenergic receptors 1 (ADRB1) and 2 (ADRB2) have not been clarified yet in detail. We, therefore, tested TAAR1 agonists TYR, PEA and OA regarding their effects on ADRB1/2 signaling by co-stimulation studies. Surprisingly, trace amines TYR and PEA are partial allosteric antagonists at ADRB1/2, whereas OA is a partial orthosteric ADRB2-antagonist and ADRB1-agonist. To specify molecular reasons for TAAR1 ligand promiscuity and for observed differences in signaling effects on particular aminergic receptors we compared TAAR, tyramine (TAR) octopamine (OAR), ADRB1/2 and dopamine receptors at the structural level. We found especially for TAAR1 that the remarkable ligand promiscuity is likely based on high amino acid similarity in the ligand-binding region compared with further aminergic receptors. On the other hand few TAAR specific properties in the ligand-binding site might determine differences in ligand-induced effects compared to ADRB1/2. Taken together, this study points to molecular details of TAAR1-ligand promiscuity and identified specific trace amines as allosteric or orthosteric ligands of particular β-adrenergic receptor subtypes

A study of phase separation in peptide-loaded HPMC films usingTzero-modulated temperature DSC, atomic force microscopy, and scanning electron microscopy

Despite the widespread use of drug-loaded polymeric systems, there is still considerable uncertainty with regard to the nature of the distribution of the drug within the polymer matrix. The aim of this investigation was to develop thermal and microscopic techniques whereby the miscibility and spatial distribution of a model peptide, cyclosporin A (CyA), in hydroxypropyl methylcellulose (HPMC) films may be studied. The new technique of Tzero-modulated temperature differential scanning calorimetry (Tzero MTDSC), scanning electron microscopy (SEM), and pulse force mode atomic force microscopy (PFM-AFM) were used in conjunction to study films prepared using a solvent evaporation process, with a solvent extraction study performed to elucidate the nature of the observed phases. Tzero MTDSC studies showed glass transitions for both the HPMC and CycA, with the Tg for the HPMC and CycA seen for the mixed systems. SEM showed two spherical phases of differing electron density. PFM-AFM also showed spheres of differing adhesion that increased in size on addition of drug. Pixel intensity analysis indicated that the smaller spheres corresponded to CycA. Exposure of the films to dichloromethane, in which CycA is soluble but HPMC is not, resulted in the presence of voids that corresponded well to the spheres suggested to correspond to the drug. It was concluded that the system had undergone extensive or complete phase separation, and that the thermal and microscopic techniques outlined above are an effective means by which this issue may be studied

Functional Uncoupling of Adenosine A 2A Receptors and Reduced Response to Caffeine in Mice Lacking Dopamine D 2 Receptors

Dopamine D(2) receptors (Rs) and adenosine A(2A)Rs are coexpressed on striatopallidal neurons, where they mediate opposing actions. In agreement with the idea that D(2)Rs tonically inhibit GABA release from these neurons, stimulation-evoked GABA release was significantly greater from striatal/pallidal slices from D(2)R null mutant (D(2)R(-/-)) than from wild-type (D(2)R(+/+)) mice. Release from heterozygous (D(2)R(+/-)) slices was intermediate. However, contrary to predictions that A(2A)R effects would be enhanced in D(2)R-deficient mice, the A(2A)R agonist CGS 21680 significantly increased GABA release only from D(2)R(+/+) slices. CGS 21680 modulation was observed when D(2)Rs were antagonized by raclopride, suggesting that an acute absence of D(2)Rs cannot explain the results. The lack of CGS 21680 modulation in the D(2)R-deficient mice was also not caused by a compensatory downregulation of A(2A)Rs in the striatum or globus pallidus. However, CGS 21680 significantly stimulated cAMP production only in D(2)R(+/+) striatal/pallidal slices. This functional uncoupling of A(2A)Rs in the D(2)R-deficient mice was not explained by reduced expression of G(s), G(olf), or type VI adenylyl cyclase. Locomotor activity induced by the adenosine receptor antagonist caffeine was significantly less pronounced in D(2)R(-/-) mice than in D(2)R(+/+) and D(2)R(+/-) mice, further supporting the idea that D(2)Rs are required for caffeine activation. Caffeine increased c-fos only in D(2)R(-/-) globus pallidus. The present results show that a targeted disruption of the D(2)R reduces coupling of A(2A)Rs on striatopallidal neurons and thereby responses to drugs that act on adenosine receptors. They also reinforce the ideas that D(2)Rs and A(2A)Rs are functionally opposed and that D(2)R-mediated effects normally predominate.Fil: Zahniser, Nancy R.. University of Colorado; Estados UnidosFil: Simosky, Johanna K.. University of Colorado; Estados UnidosFil: Mayfield, R. Dayne. University of Colorado; Estados UnidosFil: Negri, Cori A.. University of Colorado; Estados UnidosFil: Hanania, Taleen. University of Colorado; Estados UnidosFil: Larson, Gaynor A.. University of Colorado; Estados UnidosFil: Kelly, Michele A.. University of Oregon; Estados UnidosFil: Grandy, David K.. University of Oregon; Estados UnidosFil: Rubinstein, Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Low, Malcolm J.. University of Oregon; Estados UnidosFil: Fredholm, Bertil B.. Karolinska Huddinge Hospital. Karolinska Institutet; Sueci

Beyond Static Benchmarking: Using Experimental Manipulations to Evaluate Land Model Assumptions.

Land models are often used to simulate terrestrial responses to future environmental changes, but these models are not commonly evaluated with data from experimental manipulations. Results from experimental manipulations can identify and evaluate model assumptions that are consistent with appropriate ecosystem responses to future environmental change. We conducted simulations using three coupled carbon-nitrogen versions of the Community Land Model (CLM, versions 4, 4.5, and-the newly developed-5), and compared the simulated response to nitrogen (N) and atmospheric carbon dioxide (CO2) enrichment with meta-analyses of observations from similar experimental manipulations. In control simulations, successive versions of CLM showed a poleward increase in gross primary productivity and an overall bias reduction, compared to FLUXNET-MTE observations. Simulations with N and CO2 enrichment demonstrate that CLM transitioned from a model that exhibited strong nitrogen limitation of the terrestrial carbon cycle (CLM4) to a model that showed greater responsiveness to elevated concentrations of CO2 in the atmosphere (CLM5). Overall, CLM5 simulations showed better agreement with observed ecosystem responses to experimental N and CO2 enrichment than previous versions of the model. These simulations also exposed shortcomings in structural assumptions and parameterizations. Specifically, no version of CLM captures changes in plant physiology, allocation, and nutrient uptake that are likely important aspects of terrestrial ecosystems' responses to environmental change. These highlight priority areas that should be addressed in future model developments. Moving forward, incorporating results from experimental manipulations into model benchmarking tools that are used to evaluate model performance will help increase confidence in terrestrial carbon cycle projections

Beyond Static Benchmarking: Using Experimental Manipulations to Evaluate Land Model Assumptions

Land models are often used to simulate terrestrial responses to future environmental changes, but these models are not commonly evaluated with data from experimental manipulations. Results from experimental manipulations can identify and evaluate model assumptions that are consistent with appropriate ecosystem responses to future environmental change. We conducted simulations using three coupled carbon-nitrogen versions of the Community Land Model (CLM, versions 4, 4.5, and-the newly developed-5), and compared the simulated response to nitrogen (N) and atmospheric carbon dioxide (CO2) enrichment with meta-analyses of observations from similar experimental manipulations. In control simulations, successive versions of CLM showed a poleward increase in gross primary productivity and an overall bias reduction, compared to FLUXNET-MTE observations. Simulations with N and CO2 enrichment demonstrate that CLM transitioned from a model that exhibited strong nitrogen limitation of the terrestrial carbon cycle (CLM4) to a model that showed greater responsiveness to elevated concentrations of CO2 in the atmosphere (CLM5). Overall, CLM5 simulations showed better agreement with observed ecosystem responses to experimental N and CO2 enrichment than previous versions of the model. These simulations also exposed shortcomings in structural assumptions and parameterizations. Specifically, no version of CLM captures changes in plant physiology, allocation, and nutrient uptake that are likely important aspects of terrestrial ecosystems' responses to environmental change. These highlight priority areas that should be addressed in future model developments. Moving forward, incorporating results from experimental manipulations into model benchmarking tools that are used to evaluate model performance will help increase confidence in terrestrial carbon cycle projections