A Signature of Attractor Dynamics in the CA3 Region of the Hippocampus

The notion of attractor networks is the leading hypothesis for how associative memories are stored and recalled. A defining anatomical feature of such networks is excitatory recurrent connections. These “attract” the firing pattern of the network to a stored pattern, even when the external input is incomplete (pattern completion). The CA3 region of the hippocampus has been postulated to be such an attractor network; however, the experimental evidence has been ambiguous, leading to the suggestion that CA3 is not an attractor network. In order to resolve this controversy and to better understand how CA3 functions, we simulated CA3 and its input structures. In our simulation, we could reproduce critical experimental results and establish the criteria for identifying attractor properties. Notably, under conditions in which there is continuous input, the output should be “attracted” to a stored pattern. However, contrary to previous expectations, as a pattern is gradually “morphed” from one stored pattern to another, a sharp transition between output patterns is not expected. The observed firing patterns of CA3 meet these criteria and can be quantitatively accounted for by our model. Notably, as morphing proceeds, the activity pattern in the dentate gyrus changes; in contrast, the activity pattern in the downstream CA3 network is attracted to a stored pattern and thus undergoes little change. We furthermore show that other aspects of the observed firing patterns can be explained by learning that occurs during behavioral testing. The CA3 thus displays both the learning and recall signatures of an attractor network. These observations, taken together with existing anatomical and behavioral evidence, make the strong case that CA3 constructs associative memories based on attractor dynamics.     

Neuronal Mechanisms Encoding Global-to-Fine Information in Inferior-Temporal Cortex

Sugase et al. found that global information is represented at the initial transient firing of a single face-responsive neuron in inferior-temporal (IT) cortex, and that finer information is represented at the subsequent sustained firing. A feed-forward model and an attractor network are conceivable models to reproduce this dynamics. The attractor network, specifically an associative memory model, is employed to elucidate the neuronal mechanisms producing the dynamics. The results obtained by computer simulations show that a state of neuronal population initially approaches to a mean state of similar memory patterns, and that it finally converges to a memory pattern. This dynamics qualitatively coincides with that of face-responsive neurons. The dynamics of a single neuron in the model also coincides with that of a single face-responsive neuron. Furthermore, we propose two physiological experiments and predict the results from our model. Both predicted results are not explainable by the feed-forward model. Therefore, if the results obtained by actual physiological experiments coincide with our predicted results, the attractor network might be the neuronal mechanisms producing the dynamics of face-responsive neurons.     

Biglycan is internalized via a chlorpromazine-sensitive route

The small leucine-rich proteoglycan biglycan (BGN) is abundantly expressed in mesenchymal tissues. Its expression level is related to the phenotypic differentiation of cells. A dysregulation in BGN expression occurs under several pathological conditions, including glomerulonephritis, mesothelioma, pancreatic cancer and a mouse model of osteoporosis. Since the extracellular concentration of BGN is regulated both by secretion and endocytosis, we performed mechanistic studies on BGN endocytosis in human skin fibroblasts in vitro, using inhibitors of different endocytic routes. Chlorpromazine, an inhibitor of the clathrin-coated pit-pathway reduced endocytosis of BGN in human skin fibroblasts by 40%, and decreased degradation of BGN by 66% Filipin, an inhibitor of the caveolae pathway, and Tyrphostin AG 1478, a specific inhibitor of EGF-receptor phosphorylation that partially inhibits endocytosis of the structurally related proteoglycan decorin, had no influence on BGN internalization and degradation. Our data indicates that the classical clathrin-mediated endocytic pathway is a major route for the internalization of BGN. Based on the differential susceptibility to pharmacological inhibition, it appears that BGN endocytosis seems to be at least in part mechanistically different from decorin uptake.     

Role of glycosaminoglycans of biglycan in BMP-2 signaling

Recently we have reported that biglycan (BGN) promotes osteoblast differentiation and that this function is due in part to its ability to positively modulate bone morphogenetic protein (BMP) functions. In this study we investigated the role of glycosaminoglycans (GAGs) of BGN in this function using in vitro and in vivo models. C2C12 myogenic cells were treated or untreated with BMP-2 alone or in combination with glycanated, partially glycanated or de-glycanated BGN, and the effects on BMP signaling and function were assessed by Smad1/5/8 phosphorylation and alkaline phosphatase (ALP) activity. Furthermore, the effect of de-glycanation of BGN on BMP-2 induced osteogenesis was investigated employing a rat mandible defect model. The defects were filled with collagen scaffolds loaded with glycanated or de-glycanated BGN alone or in combination with a sub-optimal dose of BMP-2 (subBMP). In in vitro experiments, BMP signaling and function were the greatest when BMP-2 was combined with de-glycanated BGN among the groups tested. In the rat mandible experiments, μCT analyses revealed that the newly formed bone was significantly increased only when subBMP was combined with de-glycanated BGN. The data indicate that the GAG component of BGN functions as a suppressor for the BGN-assisted BMP function.     

Transforming growth factor-beta (TGF-beta) type I receptor/ALK5-dependent activation of the GADD45beta gene mediates the induction of biglycan expression by TGF-beta

We have recently shown that induction of biglycan (BGN) expression by transforming growth factor-beta1 (TGF-beta1) required sequential activation of both Smad and p38 mitogen-activated protein kinase signaling (Ungefroren, H., Lenschow, W., Chen, W.-B., and Kalthoff, H. (2003) J. Biol. Chem. 278, 11041-11049). Here, we have analyzed the receptors through which TGF-beta1 controls expression of BGN and GADD45beta, the latter of which is postulated to link early Smad signaling to delayed activation of p38. Ectopic expression of a dominant-negative mutant of the TGF-beta type II receptor in PANC-1 cells abrogated TGF-beta-induced BGN up-regulation. Similarly, inhibition of the TGF-beta type I receptor/ALK5 with either SB431542 or by enforced stable expression of a kinase-dead mutant greatly attenuated the TGF-beta effect on both BGN and GADD45beta expression in PANC-1 and MG-63 cells. The enhancing effect of ALK5 on TGF-beta-mediated GADD45beta and BGN expression and on GADD45beta promoter activity was also dependent on its ability to activate Smad signaling, because an ALK5 mutant defective in Smad activation (TbetaRImL45) but with an otherwise functional kinase domain failed to mediate these responses. The TGF-beta/ALK5 effect on p38 activation and BGN expression was mimicked by overexpression of GADD45beta alone (in the absence of TGF-beta stimulation) and suppressed upon antisense inhibition of GADD45beta expression. These results show that TGF-beta induces BGN expression through (the Smad-activating function of) ALK5 and GADD45beta and suggest that the sensitivity of MyD118 to activation by TGF-beta, which varies between tissues, ultimately determines the strength of the TGF-beta effect on BGN.     

The three β-1,3-glucanases from Acremonium blochii strain C59 appear to be encoded by separate genes

Three exocellular β-1,3-glucanases from Acremonium blochii strain C59, BGN3.2, BGN3.3 and BGN3.4, were purified. Two, BGN3.2 and BGN3.4 appeared to act as exo-enzymes against laminarin from Laminaria digitata, while BGN3.3 displayed an endo-mode of action. The N-terminal amino acid sequence data for BGN3.2 and BGN3.4 suggested these two enzymes may be encoded by different genes. The gene encoding the BGN3.2 glucanase was fully sequenced, and its deduced amino acid sequence was similar to those for all other sequenced fungal exo-β-1,3-glucanases. This BGN3.2 gene consists of an uninterrupted ORF of 2349 bp encoding 783 amino acids possibly with two cleavage sites for the potential removal of a pre- and pro-protein, respectively. A DNA fragment encoding a portion of the BGN3.4 gene was amplified by PCR, and the nucleotide sequence of this fragment confirmed that BGN3.2 and BGN3.4 are encoded by different genes. The internal peptide sequences of BGN3.3 were not present in the amino acid sequence deduced from the BGN3.2 gene, reinforcing the view that BGN3.3 is also genetically different to BGN3.2. Genetic differences between multiple forms of fungal β-1,3-glucanases from a single fungus have not been reported previously.     

Enhancement of tenogenic differentiation of rat tendon-derived stem cells by biglycan

Biglycan (BGN) has been identified as one of the critical components of the tendon-derived stem cells (TDSCs) niche and may be related to tendon formation. However, so far, no study has demonstrated whether the soluble BGN could induce the tenogenic differentiation of TDSCs in vitro. The aim of this study was to investigate the effect of BGN on the tenogenic differentiation of TDSCs. The proliferation and tenogenic differentiation of TDSCs exposed to different concentrations of BGN (0, 50, 100, and 500 ng/ml) were determined by the live/dead cell staining assay, CCK-8 assay, quantitative real-time polymerase chain reaction (qRT-PCR), and western blot analysis. The BGN signaling pathway of TDSCs (with and without 50 ng/ml of BGN) was determined by western blot analysis and qRT-PCR analysis. At a concentration of 50 ng/ml, BGN increased the expression of the tenogenic markers THBS-4 and TNMD at both the messenger RNA (mRNA) and protein levels. Meanwhile, 50 ng/ml of BGN inhibited the expression of the chondrogenic and osteogenic markers SOX9, ACN, and RUNX2 at both the mRNA and protein levels. Moreover, BGN (50 ng/ml) affected the expression of the components of the extracellular matrix of TDSCs. Additionally, BGN activated the Smad1/5/8 pathway as indicated by an increase in phosphorylation and demonstrated by inhibition experiments. Upregulation in the gene expression of BMP-associated receptors (BMPRII, ActR-IIa, and BMPR-Ib) and Smad pathway components (Smad4 and 8) was observed. Taken together, BGN regulates tenogenic differentiation of TDSCs via BMP7/Smad1/5/8 pathway and this regulation may provide a basic insight into treating tendon injury.     

Analysis of oversulfation in biglycan chondroitin/dermatan sulfate oligosaccharides by chip-based nanoelectrospray ionization multistage mass spectrometry

Biglycan (BGN) is a small proteoglycan that consists of a protein core containing leucine-rich repeat regions and two glycosaminoglycan (GAG) chains of either chondroitin sulfate (CS) or dermatan sulfate (DS) type. The development of novel, highly efficient analytical methods for structural identification of BGN-derived CS/DS motifs, possibly implicated in biological events, is currently the focus of research. In this work, an improved analytical method based on fully automated chip-nanoelectrospray ionization (nanoESI) in conjunction with high-capacity ion trap (HCT) multistage mass spectrometry (MS) by collision-induced dissociation (CID) was for the first time applied to BGN CS/DS oligosaccharide analysis. The CS/DS chains were released from transfected 293 BGN by β-elimination. The chain was digested with AC I lyase, and the resulting mixture was purified and subsequently separated by size exclusion chromatography (SEC). Di- and tetrasaccharide fractions were pooled and characterized in detail using the developed chip-nanoESI protocol. The chip-nanoESI MS profile in the negative ion mode revealed the presence of under-, regularly, and oversulfated species in both di- and tetrasaccharide fractions. CID MS(2)-MS(3) yielded sequence patterns consistent with unusual oversulfated 4,5-Δ-GlcA(2S)-GalNAc(4S) and 4,5-Δ-GlcA(2S)-GalNAc(6S)-IdoA(2S)-GalNAc(6S) motifs.     

Biglycan and atherosclerosis: Lessons from high cardiovascular risk conditions

Atherosclerosis (ATH) is a chronic, dynamic, evolutive process involving morphological and structural subversion of artery walls, leading to the formation of atherosclerotic plaques. ATH generally initiates during the childhood, occurring as a result of a number of changes in the intima tunica and in the media of arteries. A key event occurring during the pathobiology of ATH is the accumulation of lipoproteins in the sub-intimal spaces mediated by extracellular matrix (ECM) molecules, especially by the chondroitin sulfate/dermatan sulfate (CS/DS) –containing proteoglycans (CS/DSPGs). Among them, the proteoglycan biglycan (BGN) is critically involved in the onset and progression of ATH and evidences show that BGN represents the missing link between the pro-atherogenic status induced by both traditional and non-traditional cardiovascular risk factors and the development and progression of vascular damage. In the light of these findings, the role of BGN in dyslipidemia, hypertension, cigarette smoking, diabetes, chronic kidney disease and inflammatory status is briefly analyzed and discussed in order to shed new light on the underlying mechanisms governing the association between BGN and ATH.     

A neural mechanism of synergy formation for whole body reaching

The present study proposes a computational model for the formation of whole body reaching synergy, i.e., coordinated movements of lower and upper limbs, characterized by a focal component (the hand must reach a target) and a postural component (the center of mass must remain inside the support base). The model is based on an extension of the equilibrium point hypothesis that has been called Passive Motion Paradigm (PMP), modified in order to achieve terminal attractor features and allow the integration of multiple constraints. The model is a network with terminal attractor dynamics. By simulating it in various conditions it was possible to show that it exhibits many of the spatio-temporal features found in experimental data. In particular, the motion of the center of mass appears to be synchronized with the motion of the hand and with proportional amplitude. Moreover, the joint rotation patterns can be accounted for by a single functional degree of freedom, as shown by principal component analysis. It is also suggested that recent findings in motor imagery support the idea that the PMP network may represent the motor cognitive part of synergy formation, uncontaminated by the effect of execution.     

Attractor analysis of asynchronous Boolean models of signal transduction networks

Prior work on the dynamics of Boolean networks, including analysis of the state space attractors and the basin of attraction of each attractor, has mainly focused on synchronous update of the nodes’ states. Although the simplicity of synchronous updating makes it very attractive, it fails to take into account the variety of time scales associated with different types of biological processes. Several different asynchronous update methods have been proposed to overcome this limitation, but there have not been any systematic comparisons of the dynamic behaviors displayed by the same system under different update methods. Here we fill this gap by combining theoretical analysis such as solution of scalar equations and Markov chain techniques, as well as numerical simulations to carry out a thorough comparative study on the dynamic behavior of a previously proposed Boolean model of a signal transduction network in plants. Prior evidence suggests that this network admits oscillations, but it is not known whether these oscillations are sustained. We perform an attractor analysis of this system using synchronous and three different asynchronous updating schemes both in the case of the unperturbed (wild-type) and perturbed (node-disrupted) systems. This analysis reveals that while the wild-type system possesses an update-independent fixed point, any oscillations eventually disappear unless strict constraints regarding the timing of certain processes and the initial state of the system are satisfied. Interestingly, in the case of disruption of a particular node all models lead to an extended attractor. Overall, our work provides a roadmap on how Boolean network modeling can be used as a predictive tool to uncover the dynamic patterns of a biological system under various internal and environmental perturbations.     

Lysed and disrupted Bifidobacterium bifidum BGN4 cells promote anti-inflammatory activities in lipopolysaccharide-stimulated RAW 264.7 cells

Bifidobacterium bifidum BGN4 has been shown to improve the immune system by regulating interleukin (IL)-6 in RAW 264.7 macrophage cells. In this study, the dead cells of B. bifidum BGN4 were produced by enzymatic and physical processing to enhance the inhibition properties of pro-inflammatory cytokines using lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Notably, the secretion levels of cytokines such as interleukin (IL)-1β, IL-6, IL-8, and tumor necrosis factor (TNF)-α were decreased by the cell-wall disrupted extracts compared to heat-killed cells. The result suggests that the exposed interior-surface of B. bifidum BGN4 has a potential ability to regulate the immune-responses in the gastrointestinal tract due to major substances in inside-cell wall such as peptidoglycan and teichoic acids. In conclusion, the lysed and disrupted cells from the inside out of B. bifidum BGN4 have anti-inflammatory properties as paraprobiotic agents to control chronic inflammatory related-diseases.     

Localization of PGI (biglycan, BGN) and PGII (decorin, DCN, PG-40) genes on human chromosomes Xq13-qter and 12q, respectively

The genes for PGI (biglycan, BGN) and PGII (decorin, DCN) have been assigned to human chromosomes X and 12 by Southern analysis of a panel of human-rodent somatic cell hybrid DNAs with cDNA probes for these related small proteoglycans. Regional localization of BGN to Xq13-qter and DCN to 12p12.1-qter was also obtained by examining hybrids containing spontaneous breaks or well-characterized translocations involving chromosomes X and 12. Biglycan (BGN) is a single-copy gene about 6 kb in length. Hybridization with subfragment cDNA probes suggests the presence of two copies of the decorin (DCN) gene, or related sequences, at the locus on chromosome 12, although there is no evidence for function of more than one DCN gene. Efforts to detect restriction fragment length polymorphisms with these probes were unsuccessful.     

Construction of an associative memory using unstable periodic orbits of a chaotic attractor

Unstable periodic orbits are the skeleton of a chaotic attractor. We constructed an associative memory based on the chaotic attractor of an artificial neural network, which associates input patterns to unstable periodic orbits. By processing an input, the system is driven out of the ground state to one of the pre-defined disjunctive areas of the attractor. Each of these areas is associated with a different unstable periodic orbit. We call an input pattern learned if the control mechanism keeps the system on the unstable periodic orbit during the response. Otherwise, the system relaxes back to the ground state on a chaotic trajectory. The major benefits of this memory device are its high capacity and low-energy consumption. In addition, new information can be simply added by linking a new input to a new unstable periodic orbit.     

Patterns of ongoing activity and the functional architecture of the primary visual cortex

Ongoing spontaneous activity in the cerebral cortex exhibits complex spatiotemporal patterns in the absence of sensory stimuli. To elucidate the nature of this ongoing activity, we present a theoretical treatment of two contrasting scenarios of cortical dynamics: (1) fluctuations about a single background state and (2) wandering among multiple "attractor" states, which encode a single or several stimulus features. Studying simplified network rate models of the primary visual cortex (V1), we show that the single state scenario is characterized by fast and high-dimensional Gaussian-like fluctuations, whereas in the multiple state scenario the fluctuations are slow, low dimensional, and highly non-Gaussian. Studying a more realistic model that incorporates correlations in the feed-forward input, spatially restricted cortical interactions, and an experimentally derived layout of pinwheels, we show that recent optical-imaging data of ongoing activity in V1 are consistent with the presence of either a single background state or multiple attractor states encoding many features.     

利用放射杂交克隆板进行猪FMR1等5个基因间的连锁分析

放射杂交技术在人类图谱（包括ESTs、STSs和微卫星）的构建中已证明是一种非常有效的方法。根据人类基因组X染色体上FMR1、IDS、FATE、BGN、F8A等5个基因的信息资源,用已构建的猪/仓鼠96个放射杂交细胞系分析猪染色体该5个基因间的连锁关系。结果显示：当LOD值为4时,FMR1、IDS、FATE、BGN、F8A都处于同一个连锁群内;当LOD值为5时,FMR1、IDS处于同一个连锁群,FATE和BGN在同一个连锁群内,而F8A单独处于一个连锁群中。