Tonic inhibition of accumbal spiny neurons by extrasynaptic 4 GABAA receptors modulates the actions of psychostimulants

Within the nucleus accumbens (NAc), synaptic GABAA receptors (GABAARs) mediate phasic inhibition of medium spiny neurons (MSNs) and influence behavioral responses to cocaine. We demonstrate that both dopamine D1- and D2-receptor-expressing MSNs (D-MSNs) additionally harbor extrasynaptic GABAARs incorporating α4, β, and δ subunits that mediate tonic inhibition, thereby influencing neuronal excitability. Both the selective δ-GABAAR agonist THIP and DS2, a selective positive allosteric modulator, greatly increased the tonic current of all MSNs from wild-type (WT), but not from δ−/− or α4−/− mice. Coupling dopamine and tonic inhibition, the acute activation of D1 receptors (by a selective agonist or indirectly by amphetamine) greatly enhanced tonic inhibition in D1-MSNs but not D2-MSNs. In contrast, prolonged D2 receptor activation modestly reduced the tonic conductance of D2-MSNs. Behaviorally, WT and constitutive α4−/− mice did not differ in their expression of cocaine-conditioned place preference (CPP). Importantly, however, mice with the α4 deletion specific to D1-expressing neurons (α4D1−/−) showed increased CPP. Furthermore, THIP administered systemically or directly into the NAc of WT, but not α4−/− or α4D1−/− mice, blocked cocaine enhancement of CPP. In comparison, α4D2−/− mice exhibited normal CPP, but no cocaine enhancement. In conclusion, dopamine modulation of GABAergic tonic inhibition of D1- and D2-MSNs provides an intrinsic mechanism to differentially affect their excitability in response to psychostimulants and thereby influence their ability to potentiate conditioned reward. Therefore, α4βδ GABAARs may represent a viable target for the development of novel therapeutics to better understand and influence addictive behaviors

Carbon balance assessment of a natural steppe of southern Siberia by multiple constraint approach

Steppe ecosystems represent an interesting case in which the assessment of carbon balance may be performed through a cross validation of the eddy covariance measurements against ecological inventory estimates of carbon exchanges (Ehman et al., 2002; Curtis et al., 2002). <br><br> Indeed, the widespread presence of ideal conditions for the applicability of the eddy covariance technique, as vast and homogeneous grass vegetation cover over flat terrains (Baldocchi, 2003), make steppes a suitable ground to ensure a constrain to flux estimates with independent methodological approaches. <br><br> We report about the analysis of the carbon cycle of a true steppe ecosystem in southern Siberia during the growing season of 2004 in the framework of the TCOS-Siberia project activities performed by continuous monitoring of CO<sub>2</sub> fluxes at ecosystem scale by the eddy covariance method, fortnightly samplings of phytomass, and ingrowth cores extractions for NPP assessment, and weekly measurements of heterotrophic component of soil CO<sub>2</sub> effluxes obtained by an experiment of root exclusion. <br><br> The carbon balance of the monitored natural steppe was, according to micrometeorological measurements, a sink of carbon of 151.7±36.9 g C m<sup>−2</sup>, cumulated during the growing season from May to September. This result was in agreement with the independent estimate through ecological inventory which yielded a sink of 150.1 g C m<sup>−2</sup> although this method was characterized by a large uncertainty (±130%) considering the 95% confidence interval of the estimate. Uncertainties in belowground process estimates account for a large part of the error. Thus, in particular efforts to better quantify the dynamics of root biomass (growth and turnover) have to be undertaken in order to reduce the uncertainties in the assessment of NPP. This assessment should be preferably based on the application of multiple methods, each one characterized by its own merits and flaws

Continuous monitoring of tree responses to climate change for smart forestry: a cybernetic web of trees

6openBothTrees are long-lived organisms that contribute to forest development over centuries and beyond. However, trees are vulnerable to increasing natural and anthropic disturbances. Spatially distributed, continuous data are required to predict mortality risk and impact on the fate of forest ecosystems. In order to enable monitoring over sensitive and often remote forest areas that cannot be patrolled regularly, early warning tools/platforms of mortality risk need to be established across regions. Although remote sensing tools are good at detecting change once it has occurred, early warning tools require ecophysiological information that is more easily collected from single trees on the ground. Here, we discuss the requirements for developing and implementing such a treebased platform to collect and transmit ecophysiological forest observations and environmental measurements from representative forest sites, where the goals are to identify and to monitor ecological tipping points for rapid forest decline. Long-term monitoring of forest research plots will contribute to better understanding of disturbance and the conditions that precede it. International networks of these sites will provide a regional view of susceptibility and impacts and would play an important role in ground-truthing remotely sensed data.openTognetti, Roberto; Valentini, Riccardo; Belelli Marchesini, Luca; Gianelle, Damiano; Panzacchi, Pietro; Marshall, John D.Tognetti, R.; Valentini, R.; Belelli Marchesini, L.; Gianelle, D.; Panzacchi, P.; Marshall, J.D

During postnatal development endogenous neurosteroids influence GABA-ergic neurotransmission of mouse cortical neurons

AbstractAs neuronal development progresses, GABAergic synaptic transmission undergoes a defined program of reconfiguration. For example, GABAA receptor (GABAAR)-mediated synaptic currents, (miniature inhibitory postsynaptic currents; mIPSCs), which initially exhibit a relatively slow decay phase, become progressively reduced in duration, thereby supporting the temporal resolution required for mature network activity. Here we report that during postnatal development of cortical layer 2/3 pyramidal neurons, GABAAR-mediated phasic inhibition is influenced by a resident neurosteroid tone, which wanes in the second postnatal week, resulting in the brief phasic events characteristic of mature neuronal signalling. Treatment of cortical slices with the immediate precursor of 5α-pregnan-3α-ol-20-one (5α3α), the GABAAR-inactive 5α-dihydroprogesterone, (5α-DHP), greatly prolonged the mIPSCs of P20 pyramidal neurons, demonstrating these more mature neurons retain the capacity to synthesize GABAAR-active neurosteroids, but now lack the endogenous steroid substrate. Previously, such developmental plasticity of phasic inhibition was ascribed to the expression of synaptic GABAARs incorporating the α1 subunit. However, the duration of mIPSCs recorded from L2/3 cortical neurons derived from α1 subunit deleted mice, were similarly under the developmental influence of a neurosteroid tone. In addition to principal cells, synaptic GABAARs of L2/3 interneurons were modulated by native neurosteroids in a development-dependent manner. In summary, local neurosteroids influence synaptic transmission during a crucial period of cortical neurodevelopment, findings which may be of importance for establishing normal network connectivity

An estimate of the terrestrial carbon budget of Russia using inventory-based, eddy covariance and inversion methods

We determine the carbon balance of Russia, including Ukraine, Belarus and Kazakhstan using inventory based, eddy covariance, Dynamic Global Vegetation Models (DGVM), and inversion methods. Our current best estimate of the net biosphere to atmosphere flux is -0.66 Pg C yr-1. This sink is primarily caused by forests that using two independent methods are estimated to take up -0.69 Pg C yr-1. Using inverse models yields an average net biosphere to atmosphere flux of the same value with a interannual variability of 35%. The total estimated biosphere to atmosphere flux from eddy covariance observations over a limited number of sites amounts to -1 Pg C yr-1. Fires emit 137 to 121 Tg C yr-1 using two different methods. The interannual variability of fire emissions is large, up to a factor 0.5 to 3. Smaller fluxes to the ocean and inland lakes, trade are also accounted for. Our best estimate for the Russian net biosphere to atmosphere flux then amounts to -659 Tg C yr-1 as the average of the inverse models of -653 Tg C yr-1, bottom up -563 Tg C yr-1 and the independent landscape approach of -761 Tg C yr-1. These three methods agree well within their error bounds, so there is good consistency between bottom up and top down methods. The best estimate of the net land to atmosphere flux, including the fossil fuel emissions is -145 to -73 Tg C yr-1. Estimated methane emissions vary considerably with one inventory-based estimate providing a net land to atmosphere flux of 12.6 Tg C-CH4yr-1 and an independent model estimate for the boreal and Arctic zones of Eurasia of 27.6 Tg C-CH4yr-1

Early-life adversity selectively impairs α2-GABAA receptor expression in the mouse nucleus accumbens and influences the behavioral effects of cocaine

Haplotypes of the Gabra2 gene encoding the α2 subunit of the GABAA receptor (GABAAR) are associated with drug abuse, suggesting that α2-GABAARs may play an important role in the circuitry underlying drug misuse. The genetic association of Gabra2 haplotypes with cocaine addiction appears to be evident primarily in individuals who had experienced childhood trauma. Given this association of childhood trauma, cocaine abuse and the Gabra2 haplotypes, we have explored in a mouse model of early life adversity (ELA) whether such events influence the behavioral effects of cocaine and if, as suggested by the human studies, α2-GABAARs in the nucleus accumbens (NAc) are involved in these perturbed behaviors. In adult mice prior ELA caused a selective decrease of accumbal α2-subunit mRNA, resulting in a selective decrease in the number and size of the α2-subunit (but not the α1-subunit) immunoreactive clusters in NAc core medium spiny neurons (MSNs). Functionally, in adult MSNs ELA decreased the amplitude and frequency of GABAAR-mediated miniature inhibitory postsynaptic currents (mIPSCs), a profile similar to that of α2 "knock-out" (α2-/-) mice. Behaviorally, adult male ELA and α2-/- mice exhibited an enhanced locomotor response to acute cocaine and blunted sensitization upon repeated cocaine administration, when compared to their appropriate controls. Collectively, these findings reveal a neurobiological mechanism which may relate to the clinical observation that early trauma increases the risk for substance abuse disorder (SAD) in individuals harbouring haplotypic variations in the Gabra2 gene.</p

A pilot study comparing the metabolic profiles of elite-level athletes from different sporting disciplines

Background: The outstanding performance of an elite athlete might be associated with changes in their blood metabolic profile. The aims of this study were to compare the blood metabolic profiles between moderate- and high-power and endurance elite athletes and to identify the potential metabolic pathways underlying these differences. Methods: Metabolic profiling of serum samples from 191 elite athletes from different sports disciplines (121 high- and 70 moderate-endurance athletes, including 44 high- and 144 moderate-power athletes), who participated in national or international sports events and tested negative for doping abuse at anti-doping laboratories, was performed using non-targeted metabolomics-based mass spectroscopy combined with ultrahigh-performance liquid chromatography. Multivariate analysis was conducted using orthogonal partial least squares discriminant analysis. Differences in metabolic levels between high- and moderate-power and endurance sports were assessed by univariate linear models. Results: Out of 743 analyzed metabolites, gamma-glutamyl amino acids were significantly reduced in both high-power and high-endurance athletes compared to moderate counterparts, indicating active glutathione cycle. High-endurance athletes exhibited significant increases in the levels of several sex hormone steroids involved in testosterone and progesterone synthesis, but decreases in diacylglycerols and ecosanoids. High-power athletes had increased levels of phospholipids and xanthine metabolites compared to moderate-power counterparts. Conclusions: This pilot data provides evidence that high-power and high-endurance athletes exhibit a distinct metabolic profile that reflects steroid biosynthesis, fatty acid metabolism, oxidative stress, and energy-related metabolites. Replication studies are warranted to confirm differences in the metabolic profiles associated with athletes’ elite performance in independent data sets, aiming ultimately for deeper understanding of the underlying biochemical processes that could be utilized as biomarkers with potential therapeutic implications

Deletion of the gabra2 gene results in hypersensitivity to the acute effects of ethanol but does not alter ethanol self administration

Human genetic studies have suggested that polymorphisms of the GABRA2 gene encoding the GABA(A) α2-subunit are associated with ethanol dependence. Variations in this gene also convey sensitivity to the subjective effects of ethanol, indicating a role in mediating ethanol-related behaviours. We therefore investigated the consequences of deleting the α2-subunit on the ataxic and rewarding properties of ethanol in mice. Ataxic and sedative effects of ethanol were explored in GABA(A) α2-subunit wildtype (WT) and knockout (KO) mice using a Rotarod apparatus, wire hang and the duration of loss of righting reflex. Following training, KO mice showed shorter latencies to fall than WT littermates under ethanol (2 g/kg i.p.) in both Rotarod and wire hang tests. After administration of ethanol (3.5 g/kg i.p.), KO mice took longer to regain the righting reflex than WT mice. To ensure the acute effects are not due to the gabra2 deletion affecting pharmacokinetics, blood ethanol concentrations were measured at 20 minute intervals after acute administration (2 g/kg i.p.), and did not differ between genotypes. To investigate ethanol's rewarding properties, WT and KO mice were trained to lever press to receive increasing concentrations of ethanol on an FR4 schedule of reinforcement. Both WT and KO mice self-administered ethanol at similar rates, with no differences in the numbers of reinforcers earned. These data indicate a protective role for α2-subunits, against the acute sedative and ataxic effects of ethanol. However, no change was observed in ethanol self administration, suggesting the rewarding effects of ethanol remain unchange

Photo-antagonism of the GABAA receptor

Neurotransmitter receptor trafficking is fundamentally important for synaptic transmission and neural network activity. GABAA receptors and inhibitory synapses are vital components of brain function, yet much of our knowledge regarding receptor mobility and function at inhibitory synapses is derived indirectly from using recombinant receptors, antibody-tagged native receptors and pharmacological treatments. Here we describe the use of a set of research tools that can irreversibly bind to and affect the function of recombinant and neuronal GABAA receptors following ultraviolet photoactivation. These compounds are based on the competitive antagonist gabazine and incorporate a variety of photoactive groups. By using site-directed mutagenesis and ligand-docking studies, they reveal new areas of the GABA binding site at the interface between receptor β and α subunits. These compounds enable the selected inactivation of native GABAA receptor populations providing new insight into the function of inhibitory synapses and extrasynaptic receptors in controlling neuronal excitation