105 research outputs found
Single-molecule studies of conformational states and dynamics in the ABC importer OpuA
The current model of active transport via ABC importers is mostly based on structural, biochemical and genetic data. We here establish single-molecule Förster resonance energy transfer (smFRET) assays to monitor the conformational states and heterogeneity of the osmoregulatory type I ABC importer OpuA from Lactococcus~lactis. We present data probing both intradomain distances that elucidate conformational changes within the substrate-binding domain (SBD) OpuAC, and interdomain distances between SBDs or transmembrane domains. Using this methodology, we studied ligand-binding mechanisms, as well as ATP and glycine betaine dependences of conformational changes. Our work expands the scope of smFRET investigations towards a class of so far unstudied ABC importers, and paves the way for a full understanding of their transport cycle in the future
Cholinergic Modulation of Locomotion and Striatal Dopamine Release Is Mediated by α6α4* Nicotinic Acetylcholine Receptors
Dopamine (DA) release in striatum is governed by firing rates of midbrain DA neurons, striatal cholinergic tone, and nicotinic ACh
receptors (nAChRs) on DA presynaptic terminals. DA neurons selectively express α6* nAChRs, which show high ACh and nicotine
sensitivity. To help identify nAChR subtypes that control DA transmission, we studied transgenic mice expressing hypersensitive α6^(L9’S*)
receptors. α6^(L9’S) mice are hyperactive, travel greater distance, exhibit increased ambulatory behaviors such as walking, turning, and
rearing, and show decreased pausing, hanging, drinking, and grooming. These effects were mediated by α6 α4* pentamers, as α6^(L9’S) mice
lacking α4 subunits displayed essentially normal behavior. In α6^(L9’S) mice, receptor numbers are normal, but loss of α4 subunits leads to
fewer and less sensitive α6* receptors. Gain-of-function nicotine-stimulated DA release from striatal synaptosomes requires α4 subunits,
implicating α6α4β2* nAChRs in α6^(L9’S) mouse behaviors. In brain slices, we applied electrochemical measurements to study
control of DA release by α6^(L9’S) nAChRs. Burst stimulation of DA fibers elicited increased DA release relative to single action potentials
selectively in α6^(L9’S), but not WT or α4KO/ α6^(L9’S), mice. Thus, increased nAChR activity, like decreased activity, leads to enhanced
extracellular DA release during phasic firing. Bursts may directly enhance DA release from α6^(L9’S) presynaptic terminals, as there was no
difference in striatal DA receptor numbers or DA transporter levels or function in vitro. These results implicate α6α4β2* nAChRs in
cholinergic control of DA transmission, and strongly suggest that these receptors are candidate drug targets for disorders involving the
DA system
Orbits and phase transitions in the multifractal spectrum
We consider the one dimensional classical Ising model in a symmetric
dichotomous random field. The problem is reduced to a random iterated function
system for an effective field. The D_q-spectrum of the invariant measure of
this effective field exhibits a sharp drop of all D_q with q < 0 at some
critical strength of the random field. We introduce the concept of orbits which
naturally group the points of the support of the invariant measure. We then
show that the pointwise dimension at all points of an orbit has the same value
and calculate it for a class of periodic orbits and their so-called offshoots
as well as for generic orbits in the non-overlapping case. The sharp drop in
the D_q-spectrum is analytically explained by a drastic change of the scaling
properties of the measure near the points of a certain periodic orbit at a
critical strength of the random field which is explicitly given. A similar
drastic change near the points of a special family of periodic orbits explains
a second, hitherto unnoticed transition in the D_q-spectrum. As it turns out, a
decisive role in this mechanism is played by a specific offshoot. We
furthermore give rigorous upper and/or lower bounds on all D_q in a wide
parameter range. In most cases the numerically obtained D_q coincide with
either the upper or the lower bound. The results in this paper are relevant for
the understanding of random iterated function systems in the case of moderate
overlap in which periodic orbits with weak singularity can play a decisive
role.Comment: The article has been completely rewritten; the title has changed; a
section about the typical pointwise dimension as well as several references
and remarks about more general systems have been added; to appear in J. Phys.
A; 25 pages, 11 figures, LaTeX2
α6* Nicotinic Acetylcholine Receptor Expression and Function in a Visual Salience Circuit
Nicotinic acetylcholine receptors (nAChRs) containing α6 subunits are expressed in only a few brain areas, including midbrain dopamine (DA) neurons, noradrenergic neurons of the locus ceruleus, and retinal ganglion cells. To better understand the regional and subcellular expression pattern of α6-containing nAChRs, we created and studied transgenic mice expressing a variant α6 subunit with green fluorescent protein (GFP) fused in-frame in the M3-M4 intracellular loop. In α6-GFP transgenic mice, α6-dependent synaptosomal DA release and radioligand binding experiments confirmed correct expression and function in vivo. In addition to strong α6* nAChR expression in glutamatergic retinal axons, which terminate in superficial superior colliculus (sSC), we also found α6 subunit expression in a subset of GABAergic cell bodies in this brain area. In patch-clamp recordings from sSC neurons in brain slices from mice expressing hypersensitive α6* nAChRs, we confirmed functional, postsynaptic α6* nAChR expression. Further, sSC GABAergic neurons expressing α6* nAChRs exhibit a tonic conductance mediated by standing activation of hypersensitive α6* nAChRs by ACh. α6* nAChRs also appear in a subpopulation of SC neurons in output layers. Finally, selective activation of α6* nAChRs in vivo induced sSC neuronal activation as measured with c-Fos expression. Together, these results demonstrate that α6* nAChRs are uniquely situated to mediate cholinergic modulation of glutamate and GABA release in SC. The SC has emerged as a potential key brain area responsible for transmitting short-latency salience signals to thalamus and midbrain DA neurons, and these results suggest that α6* nAChRs may be important for nicotinic cholinergic sensitization of this pathway
How self-organized criticality works: A unified mean-field picture
We present a unified mean-field theory, based on the single site
approximation to the master-equation, for stochastic self-organized critical
models. In particular, we analyze in detail the properties of sandpile and
forest-fire (FF) models. In analogy with other non-equilibrium critical
phenomena, we identify the order parameter with the density of ``active'' sites
and the control parameters with the driving rates. Depending on the values of
the control parameters, the system is shown to reach a subcritical (absorbing)
or super-critical (active) stationary state. Criticality is analyzed in terms
of the singularities of the zero-field susceptibility. In the limit of
vanishing control parameters, the stationary state displays scaling
characteristic of self-organized criticality (SOC). We show that this limit
corresponds to the breakdown of space-time locality in the dynamical rules of
the models. We define a complete set of critical exponents, describing the
scaling of order parameter, response functions, susceptibility and correlation
length in the subcritical and supercritical states. In the subcritical state,
the response of the system to small perturbations takes place in avalanches. We
analyze their scaling behavior in relation with branching processes. In
sandpile models because of conservation laws, a critical exponents subset
displays mean-field values ( and ) in any dimensions. We
treat bulk and boundary dissipation and introduce a new critical exponent
relating dissipation and finite size effects. We present numerical simulations
that confirm our results. In the case of the forest-fire model, our approach
can distinguish between different regimes (SOC-FF and deterministic FF) studied
in the literature and determine the full spectrum of critical exponents.Comment: 21 RevTex pages, 3 figures, submitted to Phys. Rev.
Nicotinic cholinergic mechanisms causing elevated dopamine release and abnormal locomotor behavior
Firing rates of dopamine (DA) neurons in substantia nigra pars compacta (SNc) and ventral tegmental area (VTA) control DA release in target structures such as striatum and prefrontal cortex. DA neuron firing in the soma and release probability at axon terminals are tightly regulated by cholinergic transmission and nicotinic acetylcholine receptors (nAChRs). To understand the role of α6* nAChRs in DA transmission, we studied several strains of mice expressing differing levels of mutant, hypersensitive (leucine 9′ to serine [L9′S]) α6 subunits. α6 L9′S mice harboring six or more copies of the hypersensitive α6 gene exhibited spontaneous home-cage hyperactivity and novelty-induced locomotor activity, whereas mice with an equal number of WT and L9′S α6 genes had locomotor activity resembling that of control mice. α6-dependent, nicotine-stimulated locomotor activation was also more robust in high-copy α6 L9′S mice versus low-copy mice. In wheel-running experiments, results were also bi-modal; high-copy α6 L9′S animals exhibited blunted total wheel rotations during each day of a 9-day experiment, but low-copy α6 L9′S mice ran normally on the wheel. Reduced wheel running in hyperactive strains of α6 L9′S mice was attributable to a reduction in both overall running time and velocity. ACh and nicotine-stimulated DA release from striatal synaptosomes in α6 L9′S mice was well-correlated with behavioral phenotypes, supporting the hypothesis that augmented DA release mediates the altered behavior of α6 L9′S mice. This study highlights the precise control that the nicotinic cholinergic system exerts on DA transmission and provides further insights into the mechanisms and consequences of enhanced DA release
PtrWRKY19, a novel WRKY transcription factor, contributes to the regulation of pith secondary wall formation in Populus trichocarpa
WRKY proteins are one of the largest transcription factor families in higher plants and play diverse roles in various biological processes. Previous studies have shown that some WRKY members act as negative regulators of secondary cell wall formation in pith parenchyma cells. However, the regulatory mechanism of pith secondary wall formation in tree species remains largely unknown. In this study, PtrWRKY19 encoding a homolog of Arabidopsis WRKY12 was isolated from Populus trichocarpa. PtrWRKY19 was expressed in all tissues tested, with highest expression in stems, especially in pith. PtrWRKY19 was located in the nucleus and functioned as a transcriptional repressor. Ectopic expression of PtrWRKY19 in an atwrky12 mutant successfully rescued the phenotype in pith cell walls caused by the defect of AtWRKY12, suggesting that PtrWRKY19 had conserved functions for homologous AtWRKY12. Overexpression of PtrWRKY19 in poplar plants led to a significant increase in the number of pith parenchyma cells. qRT-PCR analysis showed that lignin biosynthesis-related genes were repressed in transgenic plants. In transcient reporter assays, PtrWRKY19 was identified to repress transcription from the PtoC4H2 promoter containing the conserved W-box elements. These results indicated that PtrWRKY19 may function as a negative regulator of pith secondary wall formation in poplar
Transcriptional networks for lignin biosynthesis: more complex than we thought?
Article on transcriptional networks for lignin biosynthesis
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