Hiroki Tanaka

Poly IC Triggers a Cathepsin D- and IPS-1-Dependent Pathway to Enhance Cytokine Production and Mediate Dendritic Cell Necroptosis

RIG-I-like receptors (RLRs) sense virus-derived RNA or polyinosinic-polycytidylic acid (poly IC) to exert antiviral immune responses. Here, we examine the mechanisms underlying the adjuvant effects of poly IC. Poly IC was taken up by dendritic cells (DCs), and it induced lysosomal destabilization, which, inxa0turn, activated an RLR-dependent signaling pathway. Upon poly IC stimulation, cathepsin D was released into the cytoplasm from the lysosome to interact with IPS-1, an adaptor molecule for RLRs. This interaction facilitated cathepsin D cleavage of caspase 8 and the activation of the transcription factor NF-κB, resulting in enhanced cytokine production. Further recruitment of the kinase RIP-1 to this complex initiated the necroptosis of a small number of DCs. HMGB1 released by dying cells enhanced IFN-β production in concert with poly IC. Collectively, these findings suggest that cathepsin D-triggered, IPS-1-dependent necroptosis is a mechanism that propagates the adjuvant efficacy of poly IC.

Neural Basis for Stereopsis from Second-Order Contrast Cues

Humans and animals use visual cues such as brightness and color boundaries to identify objects and navigate through environments. However, even when these cues are not available, we can effortlessly perform these tasks by using second-order cues such as contrast variation (envelope) of patterns on surfaces. Previously, numerous psychophysical studies examined properties of binocular depth processing based on the contrast-envelope cues and suggested the existence of a stereo system that uses these cues. However, its physiological substrate has not been identified yet. Here, we show that a subset of cortical neurons in cat area 18 show binocular interactions for the contrast-envelope stimuli. These neurons are capable of representing a variety of depths in the three-dimensional space based on the information available from contrast cues alone. Furthermore, these neurons show similar disparity-tuning curves for borders defined by both luminance and contrast cues. This cue-invariant tuning is consistent with a linear binocular convergence model for monocular luminance and contrast-envelope processing pathways.

Surround Suppression of V1 Neurons Mediates Orientation-Based Representation of High-Order Visual Features

Neurons with surround suppression have been implicated in processing high-order visual features such as contrast- or texture-defined boundaries and subjective contours. However, little is known regarding how these neurons encode high-order visual information in a systematic manner as a population. To address this issue, we have measured detailed spatial structures of classical center and suppressive surround regions of receptive fields of primary visual cortex (V1) neurons and examined how a population of such neurons allow encoding of various high-order features and shapes in visual scenes. Using a novel method to reconstruct structures, we found that the center and surround regions are often both elongated parallel to each other, reminiscent of on and off subregions of simple cells without surround suppression. These structures allow V1 neurons to extract high-order contours of various orientations and spatial frequencies, with a variety of optimal values across neurons. The results show that a wide range of orientations and widths of the high-order features are systematically represented by the population of V1 neurons with surround suppression.

C5-Symmetric Chiral Corannulenes: Desymmetrization of Bowl Inversion Equilibrium via “Intramolecular” Hydrogen-Bonding Network

Because of a rapid conformational inversion, bowl-shaped C5-symmetric corannulenes, though geometrically chiral, have not been directly resolved into their enantiomers. However, if this inversion equilibrium can be desymmetrized, chiral corannulenes enriched in either enantiomer can be obtained. We demonstrated this possibility using pentasubstituted corannulenes 4 and 5 carrying amide-appended thioalkyl side chains. Compound 4 displays chiroptical activity in a chiral hydrocarbon such as limonene. Because compound 5 carries a chiral center in the side chains, its enantiomers 5R and 5S show chiroptical activity even in achiral solvents such as CHCl3 and methylcyclohexane. In sharp contrast, when the side chains bear no amide functionality (1 and 2R), no chiroptical activity emerges even in limonene or with a chiral center in the side chains. Detailed investigations revealed that the peripheral amide units in 4 and 5 are hydrogen-bonded only intramolecularly along the corannulene periphery, affording cyclic amide networks with clockwise and anticlockwise geometries. Although this networking gives rise to four stereoisomers, only two, which are enantiomeric to one another, are suggested computationally to exist in the equilibrated system. In a chiral environment (chiral solvent or side chain), their thermodynamic stabilities are certainly unequal, so the bowl-inversion equilibrium can be desymmetrized. However, this is not the case when the system contains a protic solvent that can deteriorate the hydrogen-bonding network. When the enantiomeric purity of limonene as the solvent is varied, the chiroptical activity of the corannulene core changes nonlinearly with its enantiomeric excess (majority rule).

Higher-Order Architecture of Cell Adhesion Mediated by Polymorphic Synaptic Adhesion Molecules Neurexin and Neuroligin

Polymorphic adhesion molecules neurexin and neuroligin (NL) mediate asymmetric trans-synaptic adhesion, which is crucial for synapse development and function. It is not known whether or how individual synapse function is controlled by the interactions between variants and isoforms of these molecules with differing ectodomain regions. At a physiological concentration of Ca(2+), the ectodomain complex of neurexin-1 β isoform (Nrx1β) and NL1 spontaneously assembled into crystals of a lateral sheet-like superstructure topologically compatible with transcellular adhesion. Correlative light-electron microscopy confirmed extracellular sheet formation at the junctions between Nrx1β- and NL1-expressing non-neuronal cells, mimicking the close, parallel synaptic membrane apposition. The same NL1-expressing cells, however, did not form this higher-order architecture with cells expressing the much longer neurexin-1 α isoform, suggesting a functional discrimination mechanism between synaptic contacts made by different isoforms of neurexin variants.

Structural Basis for Variant-Specific Neuroligin-Binding by α-Neurexin

Neurexins (Nrxs) are presynaptic membrane proteins with a single membrane-spanning domain that mediate asymmetric trans-synaptic cell adhesion by binding to their postsynaptic receptor neuroligins. α-Nrx has a large extracellular region comprised of multiple copies of laminin, neurexin, sex-hormone-binding globulin (LNS) domains and epidermal growth factor (EGF) modules, while that of β-Nrx has but a single LNS domain. It has long been known that the larger α-Nrx and the shorter β-Nrx show distinct binding behaviors toward different isoforms/variants of neuroligins, although the underlying mechanism has yet to be elucidated. Here, we describe the crystal structure of a fragment corresponding to the C-terminal one-third of the Nrx1α ectodomain, consisting of LNS5-EGF3-LNS6. The 2.3 Å-resolution structure revealed the presence of a domain configuration that was rigidified by inter-domain contacts, as opposed to the more common flexible “beads-on-a-string” arrangement. Although the neuroligin-binding site on the LNS6 domain was completely exposed, the location of the α-Nrx specific LNS5-EGF3 segment proved incompatible with the loop segment inserted in the B+ neuroligin variant, which explains the variant-specific neuroligin recognition capability observed in α-Nrx. This, combined with a low-resolution molecular envelope obtained by a single particle reconstruction performed on negatively stained full-length Nrx1α sample, allowed us to derive a structural model of the α-Nrx ectodomain. This model will help us understand not only how the large α-Nrx ectodomain is accommodated in the synaptic cleft, but also how the trans-synaptic adhesion mediated by α- and β-Nrxs could differentially affect synaptic structure and function.

Model system of bulking and flocculation in mixed culture of Sphaerotilus sp. and Pseudomonas sp. for dissolved oxygen deficiency and high loading

Abstract A mixed continuous culture system was made up as a model for bulking and flocculation phenomena of the activated sludge to study the effect of dissolved oxygen (DO) deficiency and the effect of high organic loading. The system consisted of a floc forming bacterium and a filamentous bacterium which were isolated from the activated sludge and were identified as Pseudomonas sp. and Sphaerotilus sp., respectively. Sphaerotilus sp. had potential to cause a filamentous bulking phenomenon on the activated sludge. It was observed that the filamentous microorganism showed three kinds of growth form, filamentous form, pellet form and dispersed form, and that the floc former showed two kinds of growth form, good floc form and dispersed form. In the model system, these changes of growth form of two microorganisms, which could be thought as the cause of settling characteristics changes, depended on the DO level and the dilution rate (as a substitution for organic loading). The DO level also influence the aggregative ability of each microorganism and the maximum oxygen uptake rate, QO2max, of filamentous microorganism. The proportions of both microorganisms in model system were inverted reversibly by the DO level or the dilution rate changes.

Architecture of binocular disparity processing in monkey inferior temporal cortex.

Neurons in the inferior temporal (IT) cortex respond not only to the shape, color or texture of objects, but to the horizontal positional disparity of visual features in the right and left retinal images. IT neurons with similar shape selectivity cluster in columns. In this study, we examined how IT neurons are spatially arranged in the IT according to their selectivity for binocular disparity. With a single electrode, we simultaneously recorded extracellular action potentials from a single neuron and those from background multiple neurons at the same sites or recorded multineuronal responses at successive sites along electrode penetrations, while monkeys performed a fixation task. For neurons at each recording site, effective shapes were first determined from a set of 20 shapes presented at the zero-disparity plane. The most effective shape was then presented with varying amounts of disparity. Single neuron responses and background multiunit responses recorded at the same sites showed a similar ability of disparity discrimination and tended to share the preferred disparity, suggesting that neurons with similar disparity selectivity are clustered in the IT. We estimated from sequential recordings along electrode penetrations that the size of the neuronal clusters with similar disparity selectivity was smaller than the size of clusters with similar shape selectivity.

Behavioral evidence for visual perception of 3-dimensional surface structures in monkeys.

Human subjects perceive two crossing bars, one in front of the other, when shown a cross with disparity added to its horizontal limbs, and they also perceive neon-color spreading when shown a stereoscopic Redies-Spillmann figure. It has thus been hypothesized that the human visual system follows the principle of generic image sampling in reconstructing 3-dimensional (3-D) surface structures. Here we examine whether monkeys also perceive these surface structures. The results indicate that monkeys, like humans, perceive two crossing bars and neon-color spreading and suggest that the principle of generic image sampling may also be applied to visual perception in monkeys.

Nociceptors Boost the Resolution of Fungal Osteoinflammation via the TRP Channel-CGRP-Jdp2 Axis

Candida albicans can enter skeletal tissue through axa0skin wound in an immunocompromised host or by contamination during orthopedic surgery. Such Candida osteomyelitis is accompanied by severe pain and bone destruction. It is established that nociceptor innervation occurs in skin and bone, but the mechanisms of nociceptive modulation in fungal inflammation remain unclear. In this study, we show that C.xa0albicans stimulates Nav1.8-positive nociceptors via the β-glucan receptor Dectin-1 to induce calcitonin gene-related peptide (CGRP). This induction of CGRP is independent of Bcl-10 or Malt-1 butxa0dependent on transient receptor potential cation channel subfamily V member 1 (TRPV1)/transient receptor potential cation channel subfamily A member 1 (TRPA1) ion channels. Hindpaw β-glucan injectionxa0after Nav1.8-positive nociceptor ablation or inxa0TRPV1/TRPA1 deficiency showed dramatically increased osteoinflammation accompanied by impaired CGRP production. Strikingly, CGRP suppressed β-glucan-induced inflammation and osteoclast multinucleation via direct suppression of nuclear factor-κB (NF-κB) p65 by the transcriptional repressor Jdp2 and inhibition of actin polymerization, respectively. These findings clearly suggest a role for Dectin-1-mediated sensocrine pathways in the resolution of fungal osteoinflammation.