bluuebunny/crossref_metadata_2025 huggingface dataset:
Synergistic activation of dendritic cells by combined Toll-like receptor ligation induces superior CTL responses in vivo
Authors: Warger Tobias , Osterloh Philipp , Rechtsteiner Gerd , Fassbender Melanie , Heib Valeska , Schmid Beate , Schmitt Edgar , Schild Hansjörg , Radsak Markus P.
Published: March 2006

Toll-like receptors (TLRs) are able to interact with pathogen-derived products and their signals induce the coordinated activation of innate and adaptive immune mechanisms. Dendritic cells (DCs) play a central role in these events. As the different TLRs are able to trigger MyD88/TRIF-dependent and -independent signaling pathways, we wondered if the simultaneous activation of these signaling cascades would synergize with respect to DC activation and induce superior cytotoxic T-lymphocyte (CTL) activity in vivo. We observed that indeed the combined activation of MyD88-dependent and -independent signaling induced by TLR7 and TLR3 ligands provoked a more rapid and more sustained bone marrow–derived DC (BMDC) activation with regard to the secretion of proinflammatory cytokines, like IL-6 and IL-12p70, and the expression of costimulatory molecules like CD40, CD70, and CD86. Furthermore, in the presence of combined TLR ligand–stimulated DCs, CD4+ and CD8+ T cells were insensitive toward the inhibitory effects of regulatory T cells. Most importantly, peptide-loaded BMDCs stimulated by TLR ligand combinations resulted in a marked increase of CTL effector functions in wild-type mice in vivo. Thus, our results provide evidence that unlocking the full potential of DCs by advanced activation protocols will boost their immunogenic potential and improve DC-based vaccination strategies.
G-CSF down-regulation of CXCR4 expression identified as a mechanism for mobilization of myeloid cells
Authors: Kim Hyun Kyung , De La Luz Sierra Maria , Williams Cassin Kimmel , Gulino A. Virginia , Tosato Giovanna
Published: March 2006

CXCR4 receptor expression is required for the retention of granulocyte precursors and mature neutrophils within the bone marrow, and disruption of the SDF-1/CXCR4 axis in the bone marrow results in the mobilization of myeloid lineage cells to the peripheral circulation. We report that G-CSF down-regulates CXCR4 expression in bone marrow–derived murine and human myeloid lineage cells. When exposed to G-CSF, murine Gr1+ bone marrow myeloid cells display a time-dependent reduction of cell-surface CXCR4 and respond poorly to SDF-1 in attachment and migration assays. Bone marrow–derived cells of nonmyeloid lineage display no change in surface CXCR4 expression upon exposure to G-CSF. Compared with controls, mice treated with G-CSF for mobilization of hematopoietic progenitor cells display reduced levels of CXCR4 selectively in bone marrow Gr1+ myeloid cells. Since bone marrow myeloid cells express G-CSF receptors and G-CSF rapidly reduces CXCR4 expression in purified Gr1+ cells populations, these results provide evidence that G-CSF acts directly on myeloid lineage cells to reduce CXCR4 expression. By down-regulating CXCR4 expression in bone marrow myeloid cells and attenuating their responsiveness to SDF-1, G-CSF promotes their mobilization from the bone marrow to the peripheral blood.
TNF downmodulates the function of human CD4+CD25hi T-regulatory cells
Authors: Valencia Xavier , Stephens Geoffrey , Goldbach-Mansky Raphaela , Wilson Mildred , Shevach Ethan M. , Lipsky Peter E.
Published: March 2006

CD4+CD25+ T-regulatory cells (Tregs) play an essential role in maintaining immunologic homeostasis and preventing autoimmunity. However, little is known about the exogenous factors that regulate their differentiation and function. Here, we report that TNF inhibits the suppressive function of both naturally occurring CD4+CD25+ Tregs and TGFβ1-induced CD4+CD25+ T-regulatory cells. The mechanism of this inhibition involves signaling through TNFRII that is constitutively expressed selectively on unstimulated Tregs and that is up-regulated by TNF. TNF-mediated inhibition of suppressive function is related to a decrease in FoxP3 mRNA and protein expression by the Tregs. Notably, CD4+CD25hi Tregs isolated from patients with active rheumatoid arthritis (RA) expressed reduced levels of FoxP3 mRNA and protein and poorly suppressed the proliferation and cytokine secretion of CD4+ effector T cells in vitro. Treatment with anti-TNF antibody (infliximab) increased FOXP3 mRNA and protein expression by CD4+CD25hi Tregs and restored their suppressive function. Thus, TNF has a novel action in modulating autoimmunity, by inhibiting CD4+CD25+ Treg activity. (Blood. 2006;108:253-261)
A simplified flow cytometric assay identifies children with acute lymphoblastic leukemia who have a superior clinical outcome
Authors: Coustan-Smith Elaine , Ribeiro Raul C. , Stow Patricia , Zhou Yinmei , Pui Ching-Hon , Rivera Gaston K. , Pedrosa Francisco , Campana Dario
Published: March 2006

Bone marrow normal lymphoid progenitors (CD19+, CD10+, and/or CD34+) are exquisitely sensitive to corticosteroids and other antileukemic drugs. We hypothesized that, in patients with B-lineage acute lymphoblastic leukemia (ALL), cells with this phenotype detected early in treatment should be leukemic rather than normal. We therefore developed a simple and inexpensive flow cytometric assay for such cells and prospectively applied it to bone marrow samples collected on day 19 from 380 children with B-lineage ALL. In 211 patients (55.5%), these cells represented 0.01% or more of the mononuclear cells; results correlated remarkably well with those of more complex flow cytometric and molecular minimal residual disease (MRD) evaluations. Among 84 uniformly treated children, the 10-year incidence of relapse or remission failure was 28.8% ± 7.1% (SE) for the 42 patients with 0.01% or more leukemic cells on day 19 detected by the simplified assay versus 4.8% ± 3.3% for the 42 patients with lower levels (P = .003). These assay results were the strongest predictor of outcome, even after adjustment for competing clinicobiologic variables. Thus, this new assay would enable most treatment centers to identify a high proportion of children with ALL who have an excellent early treatment response and a high likelihood of cure. (Blood. 2006;108:97-102)
SOUTHERN AMERICAN ENGLISH PERSONAL DATIVES: THE THEORETICAL SIGNIFICANCE OF DIALECTAL VARIATION
Authors: WEBELHUTH GERT , DANNENBERG CLARE J.
Published: March 2006

The Southern Double Object Construction, a regular form in Southern vernacular varieties of American English, is well attested. Among speakers of Southern Vernacular English, its use is not stigmatized, and it appears that it can be exchanged variably with its mainstream English approximate, the “self” reflexive (e.g., Sheiwent to the store to get herisome candy vs. Sheiwent to the store to get herselfisome candy). In this article, we contextualize the Southern Double Object Construction within the scope of syntactic literature on double object constructions. We contend that although syntactic theories, such as the Principles and Parameters model, can explain Southern Double Object Constructions in general terms, they overlook idiosyncratic, language-specific properties that we argue, like Fillmore, Kay, and O'Connor (1988), constitute theoretically important information essential to evaluation of a grammar.
New insights into extracellular matrix assembly and reorganization from dynamic imaging of extracellular matrix proteins in living osteoblasts
Authors: Sivakumar Pitchumani , Czirok Andras , Rongish Brenda J. , Divakara Vivek P. , Wang Yu-Ping , Dallas Sarah L.
Published: March 2006

The extracellular matrix (ECM) has been traditionally viewed as a static scaffold that supports cells and tissues. However, recent dynamic imaging studies suggest that ECM components are highly elastic and undergo continual movement and deformation. Latent transforming growth factor beta (TGFβ) binding protein-1 (LTBP1) is an ECM glycoprotein that binds latent TGFβ and regulates its availability and activity. LTBP1 initially co-distributes with fibronectin in the extracellular matrix of osteoblasts, and depends on fibronectin for its assembly. To gain further insights into the mechanisms of assembly of LTBP1 and its spatial and temporal interactions with fibronectin, we have performed dual fluorescence time-lapse imaging of these two proteins in living osteoblasts using fluorescent probes. Time-lapse movies showed surprisingly large fibril displacements associated with cellular movement as well as occasional breaking of LTBP1 or fibronectin-containing fibrils. Individual fibrils stretched to as much as 3.5 times or contracted to as much as one fourth of their original length. Motile cells appeared to actively mediate extracellular matrix assembly by adding `globules' or `packets' of matrix material onto existing fibrils. They also actively reorganized the extracellular matrix by shunting matrix material from one location to another and exchanging fibrillar material between fibrils. This cell-mediated matrix reorganization was primarily associated with the assembly and remodeling of the initial (early) matrix, whereas mature, established ECM was more stable. Displacement vector mapping showed that different matrix fibrillar networks within the same cultures can show different dynamic motion in response to cell movement and showed that the motion of fibrils was correlated with cell motion. These data suggest novel cell-mediated mechanisms for assembly and reorganization of the extracellular matrix and highlight a role for cell motility in the assembly process.
The small GTPase R-Ras regulates organization of actin and drives membrane protrusions through the activity of PLCϵ
Authors: Ada-Nguema Aude S. , Xenias Harry , Sheetz Michael P. , Keely Patricia J.
Published: March 2006

R-Ras, an atypical member of the Ras subfamily of small GTPases, enhances integrin-mediated adhesion and signaling through a poorly understood mechanism. Dynamic analysis of cell spreading by total internal reflection fluorescence (TIRF) microscopy demonstrated that active R-Ras lengthened the duration of initial membrane protrusion, and promoted the formation of a ruffling lamellipod, rich in branched actin structures and devoid of filopodia. By contrast, dominant-negative R-Ras enhanced filopodia formation. Moreover, RNA interference (RNAi) approaches demonstrated that endogenous R-Ras contributed to cell spreading. These observations suggest that R-Ras regulates membrane protrusions through organization of the actin cytoskeleton. Our results suggest that phospholipase Cϵ (PLCϵ) is a novel R-Ras effector mediating the effects of R-Ras on the actin cytoskeleton and membrane protrusion, because R-Ras was co-precipitated with PLCϵ and increased its activity. Knockdown of PLCϵ with siRNA reduced the formation of the ruffling lamellipod in R-Ras cells. Consistent with this pathway, inhibitors of PLC activity, or chelating intracellular Ca2+ abolished the ability of R-Ras to promote membrane protrusions and spreading. Overall, these data suggest that R-Ras signaling regulates the organization of the actin cytoskeleton to sustain membrane protrusion through the activity of PLCϵ.
Apical localization of ITPK1 enhances its ability to be a modifier gene product in a murine tracheal cell model of cystic fibrosis
Authors: Yang Ling , Reece Jeff , Gabriel Sherif E. , Shears Stephen B.
Published: March 2006

A new aspect of research into the pathogenesis of cystic fibrosis (CF) is a genetics-based search for `modifier genes' that may affect the severity of CF lung disease. Using an alternative, cell biological approach, we show that ITPK1 should be considered a modifier gene. ITPK1 synthesizes an intracellular signal, inositol (3,4,5,6)-tetrakisphosphate [Ins(3,4,5,6)P4]. A bio-activatable, cell-permeable analogue of Ins(3,4,5,6)P4 inhibited Ca2+-dependent secretion of Cl- from polarized monolayers of immortalized mouse tracheal epithelial cells (MTEs). Analysis by high-pressure liquid chromatography showed endogenous Ins(3,4,5,6)P4 levels in CF MTEs were approximately 60% below those in wild-type MTEs (P<0.03). This adaptation, which improves purinergic activation of Ca2+-dependent Cl- secretion in CF MTEs, was exceptionally specific; there was no effect upon the cellular levels of all the other inositol phosphate signals. Real-time PCR provided the explanation: the level of ITPK1 expression in wild-type MTEs was twice as high as that in CF MTEs (P<0.002). The biological impact of this differential gene expression is amplified by ITPK1 being concentrated at the apical membrane of MTEs, which we discovered following confocal immunofluorescence microscopy. Compartmentalization of Ins(3,4,5,6)P4 synthesis adjacent to its site of action will enhance its regulatory capacity.
Brush border spectrin is required for early endosome recycling in Drosophila
Authors: Phillips Matthew D. , Thomas Claire M.
Published: March 2006

An apical brush border is a characteristic of many mature epithelia. This dynamic structure consists of dense microvilli supported by F-actin bundles that protrude into the apical cytoplasm, where they are crosslinked by spectrin and myosin II to form the terminal web. Little is known about the terminal web, through which vesicles transit to and from the apical membrane. Analysis of mutations in βHeavy-spectrin, the Drosophila brush border spectrin, reveals that this protein is necessary for the maintenance of Rab5 endosomes in the midgut. As a consequence, an apical H+ V-ATPase that is probably responsible for lumenal acidification is lost both from the brush border and Rab5 endosomes. Epistasis tests indicate that βHeavy-spectrin is required during endocytosis after Dynamin and before Rab5-mediated endosome activities. These data are consistent with the location of spectrin in the terminal web, and suggest that this molecule is required for correct sorting decisions at the early endosome.
Characterization of a conduit system containing laminin-5 in the human thymus: a potential transport system for small molecules
Authors: Drumea-Mirancea Mihaela , Wessels Johannes T. , Müller Claudia A. , Essl Mike , Eble Johannes A. , Tolosa Eva , Koch Manuel , Reinhardt Dieter P. , Sixt Michael , Sorokin Lydia , Stierhof York-Dieter , Schwarz Heinz , Klein Gerd
Published: March 2006

T cells develop in the thymus in a highly specialized cellular and extracellular microenvironment. The basement membrane molecule, laminin-5 (LN-5), is predominantly found in the medulla of the human thymic lobules. Using high-resolution light microscopy, we show here that LN-5 is localized in a bi-membranous conduit-like structure, together with other typical basement membrane components including collagen type IV, nidogen and perlecan. Other interstitial matrix components, such as fibrillin-1 or -2, tenascin-C or fibrillar collagen types, were also associated with these structures. Three-dimensional (3D) confocal microscopy suggested a tubular structure, whereas immunoelectron and transmission electron microscopy showed that the core of these tubes contained fibrillar collagens enwrapped by the LN-5-containing membrane. These medullary conduits are surrounded by thymic epithelial cells, which in vitro were found to bind LN-5, but also fibrillin and tenascin-C. Dendritic cells were also detected in close vicinity to the conduits. Both of these stromal cell types express major histocompatibility complex (MHC) class II molecules capable of antigen presentation. The conduits are connected to blood vessels but, with an average diameter of 2 μm, they are too small to transport cells. However, evidence is provided that smaller molecules such as a 10 kDa dextran, but not large molecules (>500 kDa), can be transported in the conduits. These results clearly demonstrate that a conduit system, which is also known from secondary lymphatic organs such as lymph nodes and spleen, is present in the medulla of the human thymus, and that it might serve to transport small blood-borne molecules or chemokines to defined locations within the medulla.
CREB activation induced by mitochondrial dysfunction triggers triglyceride accumulation in 3T3-L1 preadipocytes
Authors: Vankoningsloo Sébastien , De Pauw Aurélia , Houbion Andrée , Tejerina Silvia , Demazy Catherine , de Longueville Françoise , Bertholet Vincent , Renard Patricia , Remacle José , Holvoet Paul , Raes Martine , Arnould Thierry
Published: March 2006

Several mitochondrial pathologies are characterized by lipid redistribution and microvesicular cell phenotypes resulting from triglyceride accumulation in lipid-metabolizing tissues. However, the molecular mechanisms underlying abnormal fat distribution induced by mitochondrial dysfunction remain poorly understood. In this study, we show that inhibition of respiratory complex III by antimycin A as well as inhibition of mitochondrial protein synthesis trigger the accumulation of triglyceride vesicles in 3T3-L1 fibroblasts. We also show that treatment with antimycin A triggers CREB activation in these cells. To better delineate how mitochondrial dysfunction induces triglyceride accumulation in preadipocytes, we developed a low-density DNA microarray containing 89 probes, which allows gene expression analysis for major effectors and/or markers of adipogenesis. We thus determined gene expression profiles in 3T3-L1 cells incubated with antimycin A and compared the patterns obtained with differentially expressed genes during the course of in vitro adipogenesis induced by a standard pro-adipogenic cocktail. After an 8-day treatment, a set of 39 genes was found to be differentially expressed in cells treated with antimycin A, among them CCAAT/enhancer-binding protein α (C/EBPα), C/EBP homologous protein-10 (CHOP-10), mitochondrial glycerol-3-phosphate dehydrogenase (GPDmit), and stearoyl-CoA desaturase 1 (SCD1). We also demonstrate that overexpression of two dominant negative mutants of the cAMP-response element-binding protein CREB (K-CREB and M1-CREB) and siRNA transfection, which disrupt the factor activity and expression, respectively, inhibit antimycin-A-induced triglyceride accumulation. Furthermore, CREB knockdown with siRNA also downregulates the expression of several genes that contain cAMP-response element (CRE) sites in their promoter, among them one that is potentially involved in synthesis of triglycerides such as SCD1. These results highlight a new role for CREB in the control of triglyceride metabolism during the adaptative response of preadipocytes to mitochondrial dysfunction.
Simvastatin–Amiodarone Interaction Resulting in Rhabdomyolysis, Azotemia, and Possible Hepatotoxicity
Authors: Ricaurte Basma , Guirguis Amir , Taylor Harris C , Zabriskie Don
Published: March 2006

Objective:

To describe the fifth reported instance, as of February 15, 2006, of a severe interaction between simvastatin and amiodarone and hypothesize inhibition of CYP3A4 as the major mechanism.

Case Summary:

A 72-year-old white man (178 cm, 77.2 kg) with diabetes mellitus, hyperlipidemia, hypertension, and mild azotemia was hospitalized on September 21, 2004, with thigh weakness, achiness, and dark urine for 7 days. Coronary artery bypass had been performed on July 7, 2004. Amiodarone 200 mg/day was started on July 10, and simvastatin 80 mg/day was initiated on August 13. Laboratory testing on the present admission included creatine kinase (CK) 19 620 U/L (reference range 60–224), blood urea nitrogen 50 mg/dL, creatinine 2.6 mg/dL, aspartate aminotransferase (AST) 912 U/L (30–60), alanine aminotransferase (ALT) 748 U/L (30–60), urine myoglobin 71100 μg/L (<50), and serum myoglobin 13 877 μg/L (<110). Simvastatin and amiodarone were discontinued, and the patient was hydrated with forced alkaline diuresis. Thirteen days later, his CK was 323 U/L, creatinine 1.7 mg/dL, ALT 145 U/L, and AST 37 U/L.

Discussion:

Simvastatin is metabolized primarily by CYP3A4, and amiodarone is a recognized inhibitor of this enzyme. This may, therefore, account for the presumed drug interaction.

Conclusions:

An objective causal assessment suggests that rhabdomyolysis, renal failure, and possibly hepatotoxicity were probably related to an amiodarone–simvastatin interaction.
An Internal Conical Mandrel Technique for Fracture Toughness Measurements on Nuclear Fuel Cladding
Authors: Catherine CS , Le Boulch D , Carassou S , Ramasubramanian N , Lemaignan C
Published: March 2006

An understanding of the limiting stress level for crack initiation and propagation in a fuel cladding material is a fundamental requirement for the development of water reactor clad materials. Conventional tests, in use to evaluate fracture properties, are of limited help, because they are adapted from ASTM standards designed for thick materials, which differ significantly from fuel cladding geometry (small diameter thin-walled tubing). The Internal Conical Mandrel (1CM) test described here is designed to simulate the effect of fuel pellet diametrical increase on a cladding with an existing axial throughwall crack. It consists in forcing a cone, having a tapered increase in diameter, inside the Zircaloy cladding with an initial axial crack. The aim of this work is to quantify the crack initiation and propagation criteria for fuel cladding material. The crack propagation is monitored by a video system for obtaining crack extension δa. A finite-element (FE) simulation of the ICM test is performed in order to derive J integrals. A node release technique is applied during the FE simulation for crack propagation and the J-resistance curves (J-δa) are generated. This paper presents the test methodology, the J computation validation, and results for cold-worked stress relieved Zircaloy-4 cladding at 20° and 300°C and also for Al 7050-T7651 aluminum alloy tubing at 20°C.
Crystal Structures of the Kainate Receptor GluR5 Ligand Binding Core Dimer with Novel GluR5-Selective Antagonists
Authors: Mayer Mark L. , Ghosal Alokesh , Dolman Nigel P. , Jane David E.
Published: March 2006

Glutamate receptor (GluR) ion channels mediate fast synaptic transmission in the mammalian CNS. Numerous crystallographic studies, the majority on the GluR2-subtype AMPA receptor, have revealed the structural basis for binding of subtype-specific agonists. In contrast, because there are far fewer antagonist-bound structures, the mechanisms for antagonist binding are much less well understood, particularly for kainate receptors that exist as multiple subtypes with a distinct biology encoded by theGluR5–7,KA1, andKA2genes. We describe here high-resolution crystal structures for the GluR5 ligand-binding core complex with UBP302 and UBP310, novel GluR5-selective antagonists. The crystal structures reveal the structural basis for the high selectivity for GluR5 observed in radiolabel displacement assays for the isolated ligand binding cores of the GluR2, GluR5, and GluR6 subunits and during inhibition of glutamate-activated currents in studies on full-length ion channels. The antagonists bind via a novel mechanism and do not form direct contacts with the E723 side chain as occurs in all previously solved AMPA and kainate receptor agonist and antagonist complexes. This results from a hyperextension of the ligand binding core compared with previously solved structures. As a result, in dimer assemblies, there is a 22 Å extension of the ion channel linkers in the transition from antagonist- to glutamate-bound forms. This large conformational change is substantially different from that described for AMPA receptors, was not possible to predict from previous work, and suggests that glutamate receptors are capable of much larger movements than previously thought.
Regeneration of Vestibular Otolith Afferents after Ototoxic Damage
Authors: Zakir Mridha , Dickman J. David
Published: March 2006

Regeneration of receptor cells and subsequent functional recovery after damage in the auditory and vestibular systems of many vertebrates is well known. Spontaneous regeneration of mammalian hair cells does not occur. However, recent approaches provide hope for similar restoration of hearing and balance in humans after loss. Newly regenerated hair cells receive afferent terminal contacts, yet nothing is known about how reinnervation progresses or whether regenerated afferents finally develop normal termination fields. We hypothesized that neural regeneration in the vestibular otolith system would recapitulate the topographic phenotype of afferent innervation so characteristic of normal development. We used an ototoxic agent to produce complete vestibular receptor cell loss and epithelial denervation, and then quantitatively examined afferent regeneration at discrete periods up to 1 year in otolith maculas. Here, we report that bouton, dimorph, and calyx afferents all regenerate slowly at different time epochs, through a progressive temporal sequence. Furthermore, our data suggest that both the hair cells and their innervating afferents transdifferentiate from an early form into more advanced forms during regeneration. Finally, we show that regeneration remarkably recapitulates the topographic organization of afferent macular innervation, comparable with that developed through normative morphogenesis. However, we also show that regenerated terminal morphologies were significantly less complex than normal fibers. Whether these structural fiber changes lead to alterations in afferent responsiveness is unknown. If true, adaptive plasticity in the central neural processing of motion information would be necessitated, because it is known that many vestibular-related behaviors fully recover during regeneration.
Tuning for Spatiotemporal Frequency and Speed in Directionally Selective Neurons of Macaque Striate Cortex
Authors: Priebe Nicholas J. , Lisberger Stephen G. , Movshon J. Anthony
Published: March 2006

We recorded the responses of direction-selective simple and complex cells in the primary visual cortex (V1) of anesthetized, paralyzed macaque monkeys. When studied with sine-wave gratings, almost all simple cells in V1 had responses that were separable for spatial and temporal frequency: the preferred temporal frequency did not change and preferred speed decreased as a function of the spatial frequency of the grating. As in previous recordings from the middle temporal visual area (MT), approximately one-quarter of V1 complex cells had separable responses to spatial and temporal frequency, and one-quarter were “speed tuned” in the sense that preferred speed did not change as a function of spatial frequency. Half fell between these two extremes. Reducing the contrast of the gratings caused the population of V1 complex cells to become more separable in their tuning for spatial and temporal frequency. Contrast dependence is explained by the contrast gain of the neurons, which was relatively higher for gratings that were either both of high or both of low temporal and spatial frequency. For stimuli that comprised two spatially superimposed sine-wave gratings, the preferred speeds and tuning bandwidths of V1 neurons could be predicted from the sum of the responses to the component gratings presented alone, unlike neurons in MT that showed nonlinear interactions. We conclude that spatiotemporal modulation of contrast gain creates speed tuning from separable inputs in V1 complex cells. Speed tuning in MT could be primarily inherited from V1, but processing that occurs after V1 and possibly within MT computes selective combinations of speed-tuned signals of special relevance for downstream perceptual and motor mechanisms.
Developmental Changes in NMDA Neurotoxicity Reflect Developmental Changes in Subunit Composition of NMDA Receptors
Authors: Zhou Miou , Baudry Michel
Published: March 2006

Excitotoxicity is generally studied in dissociated neurons, cultured hippocampal slices, or intact animals. However, the requirements of dissociated neurons or cultured slices to use prenatal or juvenile rats seriously limit the advantages of these systems, whereas the complexity of intact animals prevents detailed molecular investigations. In the present experiments, we studied developmental changes in NMDA neurotoxicity in acute hippocampal slices with lactate dehydrogenase (LDH) release in medium, propidium iodide (PI) uptake, and Nissl staining as markers of cell damage. Calpain-mediated spectrin degradation was used to test calpain involvement in NMDA neurotoxicity. NMDA treatment produced increased LDH release, PI uptake, and spectrin degradation in slices from juvenile rats but not adult rats. NMDA-induced changes in slices from young rats were blocked completely by the NMDA receptor antagonist (+)-5-methyl-10,11-dihydro-5H-dibenzo [a,d] cyclohepten-5,10-imine maleate (MK-801) and by the antagonists of NR2B receptor ifenprodil andR-(R,S)-α-(4-hydroxyphenyl)-β-methyl-4-(phenylmethyl)-1-piperidine propranol and were partly blocked by calpain inhibitor III but were not affected by the NR2A-specific antagonist [(R)-[(S)-1-(4-bromo-phenyl)-ethylamino]-(2,3-dioxo-1,2,3,4-tetrahydroquinoxalin-5-yl)-methyl]-phosphonic acid. NMDA-induced changes in Nissl staining were also different in slices from young and adult rats and blocked by NR2B but not NR2A antagonists. In contrast to NMDA treatment, oxygen/glucose deprivation (OGD) induced neurotoxicity in slices from both young and adult rats, although OGD-induced toxicity was attenuated by MK-801 only in slices from young rats. Our results are consistent with the idea that NMDA-mediated toxicity is caused by activation of NR2B- but not NR2A-containing NMDA receptors leading to calpain activation and that developmental changes in NMDA toxicity reflect developmental changes in NMDA receptor subunit composition.
Remodeling of Synaptic Structure in Sensory Cortical AreasIn Vivo
Authors: Majewska Ania K. , Newton Jessica R. , Sur Mriganka
Published: March 2006

Although plastic changes are known to occur in developing and adult cortex, it remains unclear whether these changes require remodeling of cortical circuitry whereby synapses are formed and eliminated or whether they rely on changes in the strength of existing synapses. To determine the structural stability of dendritic spines and axon terminalsin vivo, we chose two approaches. First, we performed time-lapse two-photon imaging of dendritic spine motility of layer 5 pyramidal neurons in juvenile [postnatal day 28 (P28)] mice in visual, auditory, and somatosensory cortices. We found that there were differences in basal rates of dendritic spine motility of the same neuron type in different cortices, with visual cortex exhibiting the least structural dynamics. Rewiring visual input into the auditory cortex at birth, however, failed to alter dendritic spine motility, suggesting that structural plasticity rates might be intrinsic to the cortical region. Second, we investigated the persistence of both the presynaptic (axon terminals) and postsynaptic (dendritic spine) structures in young adult mice (P40–P61), using chronicin vivotwo-photon imaging in different sensory areas. Both terminals and spines were relatively stable, with >80% persisting over a 3 week period in all sensory regions. Axon terminals were more stable than dendritic spines. These data suggest that changes in network function during adult learning and memory might occur through changes in the strength and efficacy of existing synapses as well as some remodeling of connectivity through the loss and gain of synapses.
Persistent Disruption of an Established Morphine Conditioned Place Preference
Authors: Milekic Maria H. , Brown Sheena D. , Castellini Claudia , Alberini Cristina M.
Published: March 2006

In human addicts, craving and relapse are frequently evoked by the recall of memories connected to a drug experience. Established memories can become labile if recalled and can then be disrupted by several interfering events and pharmacological treatments, including inhibition of protein synthesis. Thus, reactivation of mnemonic traces provides an opportunity for disrupting memories that contribute to pathological states. Here, we tested whether the memory of a drug experience can be weakened by inhibiting protein synthesis after the reactivation of its trace. We found that an established morphine conditioned place preference (mCPP) was persistently disrupted if protein synthesis was blocked by either anisomycin or cycloheximide after the representation of a conditioning session. Unlike other types of memories, an established mCPP did not become labile after contextual recall, but required the concomitant re-experience of both the conditioning context and the drug. An established mCPP was disrupted after the conditioning session if protein synthesis was blocked selectively in the hippocampus, basolateral amygdala, or nucleus accumbens but not in the ventral tegmental area. This disruption seems to be permanent, because the preference did not return after further conditioning. Thus, established memories induced by a drug of abuse can be persistently disrupted after reactivation of the conditioning experience.
Cerebellar Synaptic Defects and Abnormal Motor Behavior in Mice Lacking α- and β-Dystrobrevin
Authors: Grady R. Mark , Wozniak David F. , Ohlemiller Kevin K. , Sanes Joshua R.
Published: March 2006

The dystrobrevins (αDB and βDB) bind directly to dystrophin and are components of a transmembrane dystrophin–glycoprotein complex (DGC) that links the cytoskeleton to extracellular proteins in many tissues. We show here that αDB, βDB, and dystrophin are all concentrated at a discrete subset of inhibitory synapses on the somata and dendrites of cerebellar Purkinje cells. Dystrophin is depleted from these synapses in mice lacking both αDB and βDB, and DBs are depleted from these synapses in mice lacking dystrophin. In dystrophin mutants and αDB,βDB double mutants, the size and number of GABA receptor clusters are decreased at cerebellar inhibitory synapses, and sensorimotor behaviors that reflect cerebellar function are perturbed. Synaptic and behavioral abnormalities are minimal in mice lacking either αDB or βDB. Together, our results show that the DGC is required for proper maturation and function of a subset of inhibitory synapses, that DB is a key component of this DGC, and that interference with this DGC leads to behavioral abnormalities. We suggest that motor deficits in muscular dystrophy patients, which are their cardinal symptoms, may reflect not only peripheral derangements but also CNS defects.
Differential Projections of Excitatory and Inhibitory Dorsal Horn Interneurons Relaying Information from Group II Muscle Afferents in the Cat Spinal Cord
Authors: Bannatyne B. Anne , Edgley Stephen A. , Hammar Ingela , Jankowska Elzbieta , Maxwell David J.
Published: March 2006

Dorsal horn interneurons with input from group II muscle spindle afferents are components of networks involved in motor control. Thirteen dorsal horn interneurons with monosynaptic group II input were characterized electrophysiologically and labeled intracellularly with Neurobiotin. Their axonal projections were traced, and neurotransmitter content was established by using immunocytochemistry. Two subpopulations were identified: five interneurons had axons that contained vesicular glutamate transporter 2 and hence were glutamatergic and excitatory. Terminals of the remaining eight interneurons were immunoreactive for the glycine transporter 2 or were apposed to gephyrin but did not contain the GABA-synthesizing enzyme glutamic acid decarboxylase and were therefore glycinergic and inhibitory. Excitatory cells were located mainly in the central region of lamina IV and had relatively small somata and restricted dendritic trees. In contrast, inhibitory interneurons were located more ventrally, in lamina V and had relatively larger somata and more extensive dendritic trees. Axonal projections of the two subpopulations differed considerably. Excitatory interneurons predominantly projected ipsilaterally, whereas most inhibitory interneurons projected both ipsilaterally and contralaterally. Three inhibitory axons formed contacts with large cholinergic cells in motor nuclei, thus revealing a novel direct coupling between inhibitory dorsal horn interneurons and motoneurons. The organization of the excitatory interneurons is consistent with current knowledge of reflex pathways to motoneurons, but the existence and connections of the inhibitory subpopulation could not be predicted from previous data. Our results indicate that these latter interneurons exercise widespread inhibitory control over a variety of cell types located on both sides of the spinal cord.
Spatial Heterogeneity in Oligodendrocyte Lineage Maturation and Not Cerebral Blood Flow Predicts Fetal Ovine Periventricular White Matter Injury
Authors: Riddle Art , Ling Luo Ning , Manese Mario , Beardsley Douglas J. , Green Lisa , Rorvik Dawn A. , Kelly Katherine A. , Barlow Clyde H. , Kelly Jeffrey J. , Hohimer A. Roger , Back Stephen A.
Published: March 2006

Although periventricular white matter injury (PWMI) is the leading cause of chronic neurological disability and cerebral palsy in survivors of premature birth, the cellular-molecular mechanisms by which ischemia-reperfusion contributes to the pathogenesis of PWMI are not well defined. To define pathophysiologic relationships among ischemia, acute cerebral white matter damage, and vulnerable target populations, we used a global cerebral ischemia-reperfusion model in the instrumented 0.65 gestation fetal sheep. We developed a novel method to make repeated measurements of cerebral blood flow using fluorescently labeled microspheres to resolve the spatial heterogeneity of flowin situin three-dimensional space. Basal flow in the periventricular white matter (PVWM) was significantly lower than in the cerebral cortex. During global cerebral ischemia induced by carotid occlusion, flow to all regions was reduced by nearly 90%. Ischemia of 30 or 37 min duration generated selective graded injury to frontal and parietal PVWM, two regions of predilection for human PWMI. Injury was proportional to the duration of ischemia and increased markedly with 45 min of ischemia to extensively damage cortical and subcortical gray matter. Surprisingly, the distribution of PVWM damage was not uniform and not explained by heterogeneity in the degree of white matter ischemia. Rather, the extent of white matter damage coincided with the presence of a susceptible population of late oligodendrocyte progenitors. These data support that although ischemia is necessary to generate PWMI, the presence of susceptible populations of oligodendrocyte progenitors underlies regional predilection to injury.
Coincidence Detection of Synaptic Inputs Is Facilitated at the Distal Dendrites after Long-Term Potentiation Induction
Authors: Xu Ning-long , Ye Chang-quan , Poo Mu-ming , Zhang Xiao-hui
Published: March 2006

Two major aspects of dendritic integration, coincidence detection and temporal integration, depend critically on the spatial and temporal properties of the dendritic summation of synaptic inputs. Neuronal activity capable of inducing synaptic long-term potentiation (LTP) leads to increased linearity of the spatial summation of synchronous EPSPs. Whether such activity can also modulate the temporal summation of EPSPs is unknown. In the present study, we examined the linearity of the summation of EPSPs spaced by different time intervals in hippocampal CA1 pyramidal neurons, before and after LTP induction. We found that LTP induction resulted in an increased linearity of summation of the potentiated input with another synchronous or asynchronous input, with a striking dendritic location-specific selectivity for the timing of the summed inputs. At distal dendrites, LTP induction led to an increased linearity of summation only for EPSPs arriving within 5 ms, thus favoring the summation of coincident inputs. In contrast, LTP induction at proximal dendrites increased the linearity of summation for EPSPs arriving within a time window of >20 ms. Furthermore, for synaptic inputs at the distal dendrite, enhanced spiking output after LTP induction was observed only for coincidently summed EPSPs, suggesting facilitated coincidence detection. In contrast, for proximal inputs, enhanced spiking output after LTP induction occurred for EPSPs arriving within a broader time window of ∼20 ms, favoring temporal integration. Such dendritic location-dependent differential modulation of coincidence detection and temporal integration by neuronal activity represents a form of activity-dependent and domain-specific plasticity in the function of dendritic information processing.
A Critical Interaction between NR2B and MAGUK in l-DOPA Induced Dyskinesia
Authors: Gardoni Fabrizio , Picconi Barbara , Ghiglieri Veronica , Polli Federica , Bagetta Vincenza , Bernardi Giorgio , Cattabeni Flaminio , Di Luca Monica , Calabresi Paolo
Published: March 2006

Abnormal function of NMDA receptor has been suggested to be correlated with the pathogenesis of Parkinson’s disease (PD) as well as with the development ofl-3,4-dihydroxyphenylalanine (l-DOPA)-induced dyskinesia. Here we show that NMDA receptor NR2 subunits display specific alterations of their subcellular distribution in striata from unilateral 6-hydroxydopamine-lesioned,l-DOPA-treated dyskinetic, andl-DOPA-treated nondyskinetic rats. Dyskinetic animals have significantly higher levels of NR2A subunit in the postsynaptic compartment than all other experimental groups, whereas NR2B subunit shows a significant reduction in both dopamine-denervated and dyskinetic rats. These events are paralleled by profound modifications of NMDA receptor NR2B subunit association with interacting elements, i.e., members of the membrane-associated guanylate kinase (MAGUK) protein family postsynaptic density-95, synapse-associated protein-97 and synapse-associated protein-102. Treatment of nondyskinetic animals with a synthetic peptide (TAT2B) able to affect NR2B binding to MAGUK proteins as well as synaptic localization of this subunit in nondyskinetic rats was sufficient to induce a shift of treated rats toward a dyskinetic motor behavior. These data indicate abnormal NR2B redistribution between synaptic and extrasynaptic membranes as an important molecular disturbance of the glutamatergic synapse involved inl-DOPA-induced dyskinesia.
Visual Deprivation Reactivates Rapid Ocular Dominance Plasticity in Adult Visual Cortex
Authors: He Hai-Yan , Hodos William , Quinlan Elizabeth M.
Published: March 2006

Brief monocular deprivation (≤3 d) induces a rapid shift in the ocular dominance of binocular neurons in the juvenile rodent visual cortex but is ineffective in adults. Here, we report that persistent, rapid, juvenile-like ocular dominance plasticity can be reactivated in adult rodent visual cortex when monocular deprivation is preceded by visual deprivation. Ocular dominance shifts in visually deprived adults are caused by a rapid depression of the response to stimulation of the deprived eye, previously only reported in juveniles, and a simultaneous potentiation of the response to stimulation of the nondeprived eye. The enhanced ocular dominance plasticity induced by visual deprivation persists for days, even if binocular vision precedes monocular deprivation. Visual deprivation also induces a significant decrease in the level of GABAAreceptors relative to AMPA receptors and a return to the juvenile form of NMDA receptors in the visual cortex, two molecular changes that we propose enable the persistent reactivation of rapid ocular dominance plasticity.
Narratives from Spanish-Speaking Children with Impaired and Typical Language Development
Authors: McCabe Allyssa , Bliss Lynn S.
Published: March 2006

The personal narratives of Spanish-speaking children with typical and impaired language development were compared across several narrative features. Thirty-nine eight- to eleven-year-old children produced narratives in English and in Spanish. Children with typical language development produced longer narratives in both English and in Spanish than children with impaired development. Narratives in Spanish produced by children with typical language development contained more actions and orientation than those produced by the children with language impairment. Significant correlations between the English and Spanish narratives were obtained for number of utterances, orientations, and actions. Bilingual aspects of narration and clinical applications are presented.
Cover Picture: Spontaneous Emission Control in Micropillar Cavities Containing a Fluorescent Molecular Dye (Adv. Mater. 6/2006)
Authors: Adawi A. M. , Cadby A. , Connolly L. G. , Hung W.‐C. , Dean R. , Tahraoui A. , Fox A. M. , Cullis A. G. , Sanvitto D. , Skolnick M. S. , Lidzey D. G.
Published: March 2006

The cover depicts a micropillar microcavity containing a fluorescent organic dye, Lumogen Red, the fabrication of which is described on p. 742 by Lidzey and co‐workers. The optical confinement resulting from the low‐dimensionality of the pillar results in a number of effects, including an enhancement in spontaneous‐emission rate. This process is confirmed using time‐resolved scanning near‐field optical microscopy. Such structures are of interest in the development of new types of low‐threshold lasers and single‐photon light sources for quantum cryptography.
Effect of helical corrugations on the low Reynolds number flow in a tube
Authors: Wang C. Y.
Published: March 2006

The low Reynolds number flow in a tube with helically corrugated wavy wall is solved for the first time. The Navier–Stokes equations are perturbed using the small wavy amplitude as parameter. For given pitch λ there exists an optimum number of circumferential waves n where the resistance is minimum. For given n there exists an optimum λ where the induced axial rotation of the flow is maximum. © 2006 American Institute of Chemical Engineers AIChE J, 2006
Hand preference in unimanual and bimanual tasks and postural effect on manual laterality in captive red‐capped mangabeys (Cercocebus torquatus torquatus)
Authors: Blois‐Heulin C. , Guitton J.S. , Nedellec‐Bienvenue D. , Ropars L. , Vallet E.
Published: March 2006

Hand preference in 11 captive red‐capped mangabeys (Cercocebus torquatus torquatus) was examined under different conditions: a free situation during spontaneous food processing, three different postural conditions (brachiating, and bipedal and tripedal standing), and a situation involving bimanual processing. Generally, individual laterality was found regardless of the task and behavior involved. However, the number of monkeys with hand preferences and the strength of the preference increased with the complexity of the tasks. The monkeys exhibited a significantly higher and positive mean manual preference index (HI) when they were hanging than when they were quadrupedal or sitting. The strength of manual preference (ABS‐HI) was in turn higher when the monkeys were hanging or bipedal than when they were quadrupedal. The strength of manual preference was higher for both the bimanual and experimental tasks than for unimanual tasks and spontaneous activities. Although our sample was too small to allow us to make any generalizations concerning lateral preferences in red‐capped mangabeys, we propose some hypotheses about the influence of posture stability and task complexity. Am. J. Primatol. 68:1–16, 2006. © 2006 Wiley‐Liss, Inc.