Biomedical subjects

Masato Okada

Publications and source records attributed to Masato Okada.

70 records · Page 4Linked to original sources

PubMed2002

Attenuation of Helicobacter pylori CagA x SHP-2 signaling by interaction between CagA and C-terminal Src kinase.

Helicobacter pylori (H. pylori) is a causative agent of gastric diseases ranging from gastritis to cancer. The CagA protein is the product of the cagA gene carried among virulent H. pylori strains and is associated with severe disease outcomes, most notably gastric carcinoma. CagA is injected from the attached H. pylori into gastric epithelial cells and undergoes tyrosine phosphorylation. The phosphorylated CagA binds and activates SHP-2 phosphatase and thereby induces a growth factor-like morphological change termed the "hummingbird phenotype." In this work, we demonstrate that CagA is also capable of interacting with C-terminal Src kinase (Csk). As is the case with SHP-2, Csk selectively binds tyrosine-phosphorylated CagA via its SH2 domain. Upon complex formation, CagA stimulates Csk, which in turn inactivates the Src family of protein-tyrosine kinases. Because Src family kinases are responsible for CagA phosphorylation, an essential prerequisite of CagA.SHP-2 complex formation and subsequent induction of the hummingbird phenotype, our results indicate that CagA-Csk interaction down-regulates CagA.SHP-2 signaling by both competitively inhibiting CagA.SHP-2 complex formation and reducing levels of CagA phosphorylation. We further demonstrate that CagA.SHP-2 signaling eventually induces apoptosis in AGS cells. Our results thus indicate that CagA-Csk interaction prevents excess cell damage caused by deregulated activation of SHP-2. Attenuation of CagA activity by Csk may enable cagA-positive H. pylori to persistently infect the human stomach for decades while avoiding excess CagA toxicity to the host.

Amino Acid Sequence↗

PubMed2002

Cutting edge: Fyn is essential for tyrosine phosphorylation of Csk-binding protein/phosphoprotein associated with glycolipid-enriched microdomains in lipid rafts in resting T cells.

In resting T cells, Csk is constitutively localized in lipid rafts by virtue of interaction with a phosphorylated adaptor protein, Csk-binding protein (Cbp)/phosphoprotein associated with glycolipid-enriched microdomains, and sets an activation threshold in TCR signaling. In this study, we examined a kinase responsible for Cbp phosphorylation in T cell membrane rafts. By analyzing T cells from Fyn-/- mice, we clearly demonstrated that Fyn, but not Lck, has its kinase activity in membrane rafts, and plays a critical role in Cbp phosphorylation, Cbp-Csk interaction, and Csk kinase activity. Naive CD44(low)CD62 ligand(high) T cells were substantially reduced in Fyn-/- mice, presumably due to the inhibition of Cbp phosphorylation. Thus, Fyn mediates Cbp-Csk interaction and recruits Csk to rafts by phosphorylating Cbp. Csk recruited to rafts would then be activated and inhibit the kinase activity of Lck to keep resting T cells in a quiescent state. Our results elucidate a negative regulatory role for Fyn in proximal TCR signaling in lipid rafts.

Adaptor Proteins, Signal Transducing↗

PubMed2002

Spatial raft coalescence represents an initial step in Fc gamma R signaling.

Characterization of lipid rafts as separated membrane microdomains consist of heterogeneous proteins suggesting that lateral assembly of rafts after Ag receptor cross-linking represents the earliest signal generating process. In line with the concept, cross-linked Ag receptors have been shown to associate with detergent-insoluble raft fraction without the aid of Src family kinases. However, it has not been established whether spatial raft coalescence could also precede Src family kinase activation. In this study, we showed that spatial raft coalescence after low-affinity FcgammaR cross-linking in RAW264.7 macrophages is independent of Src family kinase activity. The lateral raft assembly was found to be ascribed to the action of ligand-binding subunits, rather than to immunoreceptor tyrosine-based activation motif-bearing signal subunits, because monomeric murine FcgammaRIIb expressed in rat basophilic leukemia cells successfully induced spatial raft reorganization after cross-linking. We also showed that extracellular and transmembrane region of FcgammaRIIb is sufficient for raft stabilization. Moreover, this receptor fragment triggers rapid calcium mobilization and linker for activation of T cells phosphorylation, in a manner sensitive to Src family kinase inhibition and to cholesterol depletion. Presence of immunoreceptor tyrosine-based inhibitory motif and addition of immunoreceptor tyrosine-based activation motif to the receptor fragment abolished and enhanced the responses, respectively, but did not affect raft stabilization. These findings support the concept that ligand-binding subunit is responsible for raft coalescence, and that this event triggers initial biochemical signaling.

Animals↗

PubMed2002

Acceleration effect of coupled oscillator systems.

We have developed a curved isochron clock (CIC) by modifying the radial isochron clock to provide a clean example of the acceleration (deceleration) effect. By analyzing a two-body system of coupled CICs, we determined that an unbalanced mutual interaction caused by curved isochron sets is the minimum mechanism needed for generating the acceleration (deceleration) effect in coupled oscillator systems. From this we can see that the Sakaguchi and Kuramoto (SK) model, which is a class of nonfrustrated mean field model, has an acceleration (deceleration) effect mechanism. To study frustrated coupled oscillator systems, we extended the SK model to two oscillator associative memory models, one with symmetric and the other with asymmetric dilution of coupling, which also have the minimum mechanism of the acceleration (deceleration) effect. We theoretically found that the Onsager reaction term (ORT), which is unique to frustrated systems, plays an important role in the acceleration (deceleration) effect. These two models are ideal for evaluating the effect of the ORT because, with the exception of the ORT, they have the same order parameter equations. We found that the two models have identical macroscopic properties, except for the acceleration effect caused by the ORT. By comparing the results of the two models, we can extract the effect of the ORT from only the rotation speeds of the oscillators.

Animals↗

PubMed2002

Structure of the carboxyl-terminal Src kinase, Csk.

The carboxyl-terminal Src kinase (Csk) is an indispensable negative regulator for the Src family tyrosine kinases (SFKs) that play pivotal roles in various cell signalings. To understand the molecular basis of the Csk-mediated regulation of SFKs, we elucidated the crystal structure of full-length Csk. The Csk crystal consists of six molecules classified as active or inactive states according to the coordinations of catalytic residues. Csk assembles the SH2 and SH3 domains differently from inactive SFKs, and their binding pockets are oriented outward enabling the intermolecular interaction. In active molecules, the SH2-kinase and SH2-SH3 linkers are tightly stuck to the N-lobe of the kinase domain to stabilize the active conformation, and there is a direct linkage between the SH2 and the kinase domains. In inactive molecules, the SH2 domains are rotated destroying the linkage to the kinase domain. Cross-correlation matrices for the active molecules reveal that the SH2 domain and the N-lobe of the kinase domain move as a unit. These observations suggest that Csk can be regulated through coupling of the SH2 and kinase domains and that Csk provides a novel built-in activation mechanism for cytoplasmic tyrosine kinases.

Amino Acid Sequence↗

PubMed2002

Cutting Edge: Transmembrane phosphoprotein Csk-binding protein/phosphoprotein associated with glycosphingolipid-enriched microdomains as a negative feedback regulator of mast cell signaling through the FcepsilonRI.

Tyrosine phosphorylation in the cytoplasmic domains of FcepsilonRI by the Src family kinase Lyn initiates a signaling cascade leading to mast cell activation. In this study, we show that a recently identified transmembrane protein, Csk-binding protein (Cbp), also known as phospoprotein associated with glycosphingolipid-enriched microdomains (PAG), negatively regulates FcepsilonRI signaling. In rat basophilic leukemia (RBL)-2H3 cells, the levels of tyrosine phosphorylation of Cbp/PAG and its association with Csk, a negative regulator for Lyn, significantly elevate immediately after aggregation of FcepsilonRI. An overexpression of Cbp/PAG in RBL-2H3 cells inhibits FcepsilonRI-mediated cell activation. This is accompanied with decreased levels of tyrosine phosphorylation of FcepsilonRI, association of FcepsilonRI with Lyn, and FcepsilonRI-associated tyrosine kinase activity. These findings combined with the fact that Cbp/PAG, Lyn, and aggregated FcepsilonRI are localized to lipid rafts, suggest that upon FcepsilonRI aggregation Cbp/PAG down-regulates the receptor-associated Lyn activity through relocating Csk to rafts, thereby efficiently mediating feedback inhibition of FcepsilonRI signaling.

Animals↗

PubMed2002

Cutting edge: negative regulation of immune synapse formation by anchoring lipid raft to cytoskeleton through Cbp-EBP50-ERM assembly.

Ag recognition by T lymphocytes induces immune synapse formation and recruitment of signaling molecules into a lipid raft. Cbp/PAG is a Csk-associated membrane adapter protein exclusively localized in a lipid raft. We identified NHERF/EBP50 as a Cbp-binding molecule, which connects the membrane raft and cytoskeleton by binding to both Cbp through its PDZ domain and ezrin-radixin-moesin through the C terminus. Overexpression of Cbp reduced the mobility of the raft on the cell surface of unstimulated T cells and prevented synapse formation and subsequent T cell activation, whereas a mutant incapable of EBP50 binding restored both synapse formation and activation. These results suggest that anchoring of lipid raft to the cytoskeleton through Cbp-EBP50-ezrin-radixin-moesin assembly regulates membrane dynamism for synapse formation and T cell activation.

Adaptor Proteins, Signal Transducing↗

PubMed2002

Csk, a critical link of g protein signals to actin cytoskeletal reorganization.

Heterotrimeric G proteins can signal to reorganize the actin cytoskeleton, but the mechanism is unclear. Here we report that, in tyrosine kinase Csk-deficient mouse embryonic fibroblast cells, G protein (Gbetagamma, Galpha(12), Galpha(13), and Galpha(q))-induced, and G protein-coupled receptor-induced, actin stress fiber formation was completely blocked. Reintroduction of Csk into Csk-deficent cells restored the G protein-induced actin stress fiber formation. Chemical rescue experiments with catalytic mutants of Csk demonstrated that the catalytic activity of Csk was required for this process. Furthermore, we uncovered that Gbetagamma can both translocate Csk to the plasma membrane and directly increase Csk kinase activity. Our genetic and biochemical studies demonstrate that Csk plays a critical role in mediating G protein signals to actin cytoskeletal reorganization.

3T3 Cells↗

PubMed2002

Self-regulation mechanism of temporally asymmetric Hebbian plasticity.

Recent biological experimental findings have shown that synaptic plasticity depends on the relative timing of the pre- and postsynaptic spikes. This determines whether long-term potentiation (LTP) or long-term depression (LTD) is induced. This synaptic plasticity has been called temporally asymmetric Hebbian plasticity (TAH). Many authors have numerically demonstrated that neural networks are capable of storing spatiotemporal patterns. However, the mathematical mechanism of the storage of spatiotemporal patterns is still unknown, and the effect of LTD is particularly unknown. In this article, we employ a simple neural network model and show that interference between LTP and LTD disappears in a sparse coding scheme. On the other hand, the covariance learning rule is known to be indispensable for the storage of sparse patterns. We also show that TAH has the same qualitative effect as the covariance rule when spatiotemporal patterns are embedded in the network.

Animals↗

PubMed2001

Multibranch entrainment and slow evolution among branches in coupled oscillators.

In globally coupled oscillators, it is believed that strong higher harmonics of coupling functions are essential for multibranch entrainment (MBE), in which there exist many stable states, whose number scales as approximately O(expN) (where N is the system size). The existence of MBE implies the nonergodicity of the system. Then, because this apparent breaking of ergodicity is caused by microscopic energy barriers, this seems to be in conflict with a basic principle of statistical physics. Using macroscopic dynamical theories, we demonstrate that there is no such ergodicity breaking, and such a system slowly evolves among branch states, jumping over microscopic energy barriers due to the influence of thermal noise. This phenomenon can be regarded as an example of slow dynamics driven by a perturbation along a neutrally stable manifold consisting of an infinite number of branch states.

Journal Article↗

PubMed2000

Clinical study of surgical treatment of carpal tunnel syndrome: Open versus endoscopic technique.

Twenty-six patients (30 hands) who underwent endoscopic carpal tunnel release were physically examined and asked to complete questionnaires on their symptoms, functional limitations and satisfaction. Their mean grip strength improved considerably from 17.5 kg before the operation to 31.3 kg at the final follow-up. Symptoms at the final follow-up were hypesthesia in 9 hands, muscle atrophy in 10, positive Tinel sign in 3, and positive Phalen sign in 1. The mean symptom severity score improved from 3.01 before the operation to 1.44 at the final follow-up and the mean functional status score improved from 3.20 to 1.54. The average times to return to activities of daily living and work were 7.3 days and 18.4 days respectively. Twenty-two of the 26 patients (85%) were satisfied. There was no injury to neurovascular structures. Arterial injuries were prevented by marking the superficial palmar arch and ulnar artery with a Doppler ultrasonic hemodrometer before surgery.

Journal Article↗

PubMed1997

Neural Network Model of Spatial Memory: Associative Recall of Maps.

This paper offers a neural network model that can memorize and recall spatial maps. When driving through a place we have been before, we can recall and imagine the scenery that we cannot see yet but shall see soon. Triggered by the newly recalled image, we can also recall other scenery further ahead of us. The model emulates such a chain process of recalling using a correlation matrix memory. A correlation matrix memory by itself, however, does not accept shifts in location of stimulus patterns, and each stimulus pattern has to be placed accurately at the location of one of the memorized patterns. We propose adjusting the location of the stimulus pattern using the cross-correlation between the stimulus pattern and the "piled pattern", which is the sum of all patterns memorized in the correlation matrix. A map of Europe is divided into a number of overlapping segments, and these segments are memorized in the proposed model. Triggered by an input image, say a map around Scotland, the model can recall maps of other parts of Europe sequentially up to Italy, for example. Copyright 1997 Elsevier Science Ltd.

Journal Article↗

PubMed1996

Notions of Associative Memory and Sparse Coding.

This paper summarizes associative memory models and sparse representation of memory in these models. Important properties of the associative memory models are their storage capacity, basin of attraction, and the existence of spurious memories. Sparse coding and nonmonotonic output functions are proposed to improve them. Sparsely coded associative memory model has an extremely large storage capacity which diverges as the mean firing rate of memory patterns approaches 0. The storage capacity strongly depends on the shape of the output function as well as the mean firing rate, even in the case of monotonic output functions. Dynamical properties of the model are analyzed by means of a statistical neurodynamical method. We emphasize the necessity of a feedback mechanism to control the mean firing rate in the recall process. Recently, there have been some experimental results suggesting its existence in the brain. On the other hand, it has been shown that the storage capacity can be markedly improved by replacing the usual monotonic output function with a nonmonotonic one. Another remarkable property of the model with the nonmonotonic neurons is that it seems to have no, or almost no, spurious memory. An associative memory model using nonmonotonic modules with a feedforward inhibition is discussed. The modules consist of two types of threshold units, each of which has a different threshold and can be considered as a biologically plausible representation of the nonmonotonic output function. The above model is compared with the monotonic one. The difference in the storage capacity between the two models becomes small when the sparse patterns are stored. Finally, we discuss the biological plausibility of the discussed associative memory models and sparse coding. Copyright 1996 Elsevier Science Ltd.

Journal Article↗

PubMedDate not supplied

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.

Face↗

PubMedDate not supplied

Self-organization of globally continuous and locally distributed information representation.

A number of findings suggest that the preferences of neighboring neurons in the inferior temporal (IT) cortex of macaque monkeys tend to be similar. However, a recent study reports convincingly that the preferences of neighboring neurons actually differ. These findings seem contradictory. To explain this conflict, we propose a new view of information representation in the IT cortex. This view takes into account sparse and local neuronal excitation. Since the excitation is sparse, information regarding visual objects seems to be encoded in a distributed manner. The local excitation of neurons coincides with the classical notion of a column structure. Our model consists of input layer and output layer. The main difference from conventional models is that the output layer has local and random intra-layer connections. In this paper, we adopt two rings embedded in three-dimensional space as an input signal space, and examine how resultant information representation depends on the distance between two rings that is denoted as D. We show that there exists critical value for the distance Dc. When D > Dc the output layer becomes able to form the column structure, this model can obtain the distributed representation within the column. While the output layer acquires the conventional information representation observed in the V1 cortex when D < Dc. Moreover, we consider the origin of the difference between information representation of the V1 cortex and that of the IT cortex. Our finding suggests that the difference in the information representations between the V1 and the IT cortices could be caused by difference between the input space structures.

Animals↗

PubMedDate not supplied

Regulation of T cell-dependent autoantibody production by a gammadelta T cell line derived from lupus-prone mice.

Lupus-prone (MRLxC57BL/6) F(1) mice lacking gammadelta T cells show more severe lupus than their T cell-intact counterparts, suggesting that gammadelta T cells down-modulate murine lupus. To determine the mechanisms for this effect, we assessed the capacity of gammadelta T cell lines derived from spleens of alphabeta T cell-deficient MRL/Mp-Fas(lpr) (MRL/Fas(lpr)) mice to down-regulate anti-dsDNA production generated by CD4(+)alphabeta T helper cell lines and activated B cells from wild-type MRL/Fas(lpr) mice. One line, GD12 (gd TCR(+), CD4(-)CD8(-)), had the capacity to reduce anti-dsDNA production in a contact-dependent manner. GD12 also killed activated MRL/Fas(lpr) (H-2(k)) B cells, with less cytolysis of resting B cells than that generated by in comparison to cytokine-matched gammadelta T cell lines. In addition, GD12 also killed activated B cells derived from C57BL/6-Fas(lpr) (H-2(b)) or beta(2)-microglobulin (beta(2) M)-deficient MRL/Fas(lpr) mice, suggesting cytolysis was neither MHC- nor CD1-restricted. Killing by GD12 was inhibited by anti-TNFalpha and anti-TNF-R1, and partially blocked by anti-gd TCR Fab fragments, but not by anti-FasL, anti-TNF-R2 (p75) or concanamycin A. IL-10 produced by GD12 also partially inhibited alphabeta Th1-dependent but not alphabeta Th2-dependent autoantibody production. These findings prove that we have identtified a gammadelta T cell line that suppresses autoantibody synthesis by alphabeta T-B cell collaboration in vitro.

Animals↗