Mechanical activation opens a lipid-lined pore in OSCA ion channels.

OSCA/TMEM63 channels are the largest known family of mechanosensitive channels1-3, playing critical roles in plant4-7 and mammalian8,9 mechanotransduction. Here we determined 44 cryogenic electron microscopy structures of OSCA/TMEM63 channels in different environments to investigate the molecular basis of OSCA/TMEM63 channel mechanosensitivity. In nanodiscs, we mimicked increased membrane tension and observed a dilated pore with membrane access in one of the OSCA1.2 subunits. In liposomes, we captured the fully open structure of OSCA1.2 in the inside-in orientation, in which the pore shows a large lateral opening to the membrane. Unusually for ion channels, structural, functional and computational evidence supports the existence of a 'proteo-lipidic pore' in which lipids act as a wall of the ion permeation pathway. In the less tension-sensitive homologue OSCA3.1, we identified an 'interlocking' lipid tightly bound in the central cleft, keeping the channel closed. Mutation of the lipid-coordinating residues induced OSCA3.1 activation, revealing a conserved open conformation of OSCA channels. Our structures provide a global picture of the OSCA channel gating cycle, uncover the importance of bound lipids and show that each subunit can open independently. This expands both our understanding of channel-mediated mechanotransduction and channel pore formation, with important mechanistic implications for the TMEM16 and TMC protein families.

Near-Infrared Photothermal Manipulates Cellular Excitability and Animal Behavior in Caenorhabditis elegans.

Near-infrared (NIR) photothermal manipulation has emerged as a promising and noninvasive technology for neuroscience research and disease therapy for its deep tissue penetration. NIR stimulated techniques have been used to modulate neural activity. However, due to the lack of suitable in vivo control systems, most studies are limited to the cellular level. Here, a NIR photothermal technique is developed to modulate cellular excitability and animal behaviors in Caenorhabditis elegans in vivo via the thermosensitive transient receptor potential vanilloid 1 (TRPV1) channel with an FDA-approved photothermal agent indocyanine green (ICG). Upon NIR stimuli, exogenous expression of TRPV1 in AFD sensory neurons causes Ca2+ influx, leading to increased neural excitability and reversal behaviors, in the presence of ICG. The GABAergic D-class motor neurons can also be activated by NIR irradiation, resulting in slower thrashing behaviors. Moreover, the photothermal manipulation is successfully applied in different types of muscle cells (striated muscles and nonstriated muscles), enhancing muscular excitability, causing muscle contractions and behavior changes in vivo. Altogether, this study demonstrates a noninvasive method to precisely regulate the excitability of different types of cells and related behaviors in vivo by NIR photothermal manipulation, which may be applied in mammals and clinical therapy.

Bone serves as a transfer station for secondary dissemination of breast cancer.

Metastasis is responsible for the majority of deaths among breast cancer patients. Although parallel polyclonal seeding has been shown to contribute to organ-specific metastasis, in the past decade, horizontal cross-metastatic seeding (metastasis-to-metastasis spreading) has also been demonstrated as a pattern of distant metastasis to multiple sites. Bone, as the most frequent first destination of breast cancer metastasis, has been demonstrated to facilitate the secondary dissemination of breast cancer cells. In this review, we summarize the clinical and experimental evidence that bone is a transfer station for the secondary dissemination of breast cancer. We also discuss the regulatory mechanisms of the bone microenvironment in secondary seeding of breast cancer, focusing on stemness regulation, quiescence-proliferation equilibrium regulation, epigenetic reprogramming and immune escape of cancer cells. Furthermore, we highlight future research perspectives and strategies for preventing secondary dissemination from bone.

Extracellular Vesicle-Packaged CDH11 and ITGA5 Induce the Premetastatic Niche for Bone Colonization of Breast Cancer Cells.

Although most breast cancer metastases in bone cause osteolytic lesions, the osteogenic niche has commonly been described as an initiator of early-stage bone colonization of disseminated cancer cells. Tumor cell-derived extracellular vesicles (EV) have been shown to determine the organotropism of cancer cells by transferring their cargo, such as nucleic acids and proteins, to resident cells at future metastatic sites and preparing a favorable premetastatic niche. Runt-related transcription factor 2 (RUNX2) and its regulated genes have been shown to facilitate the acquisition of osteomimetic features and to enhance the bone metastatic potential of breast cancer cells. In this study, we present in vivo and in vitro evidence to clarify the role of EVs released by breast cancer cells with high RUNX2 expression in the education of osteoblasts to form an osteogenic premetastatic niche. Furthermore, different extracellular vesicular proteins were identified that mediate events subsequent to the specific recognition of tumor-derived EVs by osteoblasts via cadherin 11 (CDH11) and the induction of the osteogenic premetastatic niche by integrin α5 (ITGA5). CDH11high/ITGA5high EVs were demonstrated to be responsible for the formation of a premetastatic niche that facilitates RUNX2 high-expressing breast cancer cell colonization in bone, revealing a potential EV-based premetastatic niche blockage strategy. SIGNIFICANCE: This study provides mechanistic insights into the generation of an osteogenic premetastatic niche by breast cancer-derived EVs and identifies potential EV-derived diagnostic biomarkers and targets for breast cancer bone metastasis.

Curvilinear features are important for animate/inanimate categorization in macaques.

The current experiment investigated the extent to which perceptual categorization of animacy (i.e., the ability to discriminate animate and inanimate objects) is facilitated by image-based features that distinguish the two object categories. We show that, with nominal training, naïve macaques could classify a trial-unique set of 1000 novel images with high accuracy. To test whether image-based features that naturally differ between animate and inanimate objects, such as curvilinear and rectilinear information, contribute to the monkeys' accuracy, we created synthetic images using an algorithm that distorted the global shape of the original animate/inanimate images while maintaining their intermediate features (Portilla & Simoncelli, 2000). Performance on the synthesized images was significantly above chance and was predicted by the amount of curvilinear information in the images. Our results demonstrate that, without training, macaques can use an intermediate image feature, curvilinearity, to facilitate their categorization of animate and inanimate objects.

Sensory Glia Detect Repulsive Odorants and Drive Olfactory Adaptation.

Glia are typically considered as supporting cells for neural development and synaptic transmission. Here, we report an active role of a glia in olfactory transduction. As a polymodal sensory neuron in C. elegans, the ASH neuron is previously known to detect multiple aversive odorants. We reveal that the AMsh glia, a sheath for multiple sensory neurons including ASH, cell-autonomously respond to aversive odorants via G-protein-coupled receptors (GPCRs) distinct from those in ASH. Upon activation, the AMsh glia suppress aversive odorant-triggered avoidance and promote olfactory adaptation by inhibiting the ASH neuron via GABA signaling. Thus, we propose a novel two-receptor model where the glia and sensory neuron jointly mediate adaptive olfaction. Our study reveals a non-canonical function of glial cells in olfactory transduction, which may provide new insights into the glia-like supporting cells in mammalian sensory procession.

Curvature processing in human visual cortical areas.

Curvature is one of many visual features shown to be important for visual perception. We recently showed that curvilinear features provide sufficient information for categorizing animate vs. inanimate objects, while rectilinear features do not (Zachariou et al., 2018). Results from our fMRI study in rhesus monkeys (Yue et al., 2014) have shed light on some of the neural substrates underlying curvature processing by revealing a network of visual cortical patches with a curvature response preference. However, it is unknown whether a similar network exists in human visual cortex. Thus, the current study was designed to investigate cortical areas with a preference for curvature in the human brain using fMRI at 7T. Consistent with our monkey fMRI results, we found a network of curvature preferring cortical patches-some of which overlapped well-known face-selective areas. Moreover, principal component analysis (PCA) using all visually-responsive voxels indicated that curvilinear features of visual stimuli were associated with specific retinotopic regions in visual cortex. Regions associated with positive curvilinear PC values encompassed the central visual field representation of early visual areas and the lateral surface of temporal cortex, while those associated with negative curvilinear PC values encompassed the peripheral visual field representation of early visual areas and the medial surface of temporal cortex. Thus, we found that broad areas of curvature preference, which encompassed face-selective areas, were bound by central visual field representations. Our results support the hypothesis that curvilinearity preference interacts with central-peripheral processing biases as primary features underlying the organization of temporal cortex topography in the adult human brain.

Distinct functions of TMC channels: a comparative overview.

In the past two decades, transmembrane channel-like (TMC) proteins have attracted a significant amount of research interest, because mutations of Tmc1 lead to hereditary deafness. As evolutionarily conserved membrane proteins, TMC proteins are widely involved in diverse sensorimotor functions of many species, such as hearing, chemosensation, egg laying, and food texture detection. Interestingly, recent structural and physiological studies suggest that TMC channels may share a similar membrane topology with the Ca2+-activated Cl- channel TMEM16 and the mechanically activated OSCA1.2/TMEM63 channel. Namely, these channels form dimers and each subunit consists of ten transmembrane segments. Despite this important structural insight, a key question remains: what is the gating mechanism of TMC channels? The major technical hurdle to answer this question is that the reconstitution of TMC proteins as functional ion channels has been challenging in mammalian heterologous systems. Since TMC channels are conserved across taxa, genetic studies of TMC channels in model organisms such as C. elegans, Drosophila, and zebrafish may provide us critical information on the physiological function and regulation of TMCs. Here, we present a comparative overview on the diverse functions of TMC channels in different species.

Reusable Silicon-Based SERS Chip for Ratiometric Analysis of Fluoride Ion in Aqueous Solutions.

An innovative ratiometric surface-enhanced Raman scattering (SERS) sensor using a 4-mercaptoboric acid (4-MPBA)-modified silver nanoparticle-decorated silicon wafer (Si@Ag NPs chip) was proposed for the ultrasensitive determination of F- ions in aqueous solutions. The principle of sensing strategy is based on fluoride-induced structural symmetry breaking and charge redistribution of phenylboronic acid, leading to a band shift of the C-C stretching mode of 4-MPBA from 1589 to 1576 cm-1. Accordingly, a ratiometric signal of the area ratio (A1576/A1589) between the fluoride-bond MPBA molecules and unoccupied MPBA molecules can be used for the quantitative response of F- ions. In comparison with other SERS-based sensing methods, this ratiometric method can avoid a large error resulting from the inhomogeneity of substrates. Under the optimized analytical conditions, the proposed SERS sensor possesses a quick response to F- ions within 2 min and exhibits high selectivity for F- ions with the determination limit of 10-8 M, which is over 3 orders of magnitude lower than the World Health Organization (WHO) guideline value for F- ions in drinking water. Of particular significance, the present sensor features favorable recyclability, which preserves suitable reproducibility during 6-time cyclic determination of F- ions. The practical utility of this sensing system for the determination of F- ions was tested with real water and toothpaste samples, and the results demonstrate that this sensor shows high recoveries (90-110%). Given its simple principle and easy operation, the present silicon-based SERS sensor could serve as a promising sensor for various practical applications.

Serotonergic neuron ADF modulates avoidance behaviors by inhibiting sensory neurons in C. elegans.

Serotonin plays an essential role in both the invertebrate and vertebrate nervous systems. ADF, an amphid neuron with dual ciliated sensory endings, is considered to be the only serotonergic sensory neuron in the hermaphroditic Caenorhabditis elegans. This neuron is known to be involved in a range of behaviors including pharyngeal pumping, dauer formation, sensory transduction, and memory. However, whether ADF neuron is directly activated by environmental cues and how it processes these information remains unknown. In this study, we found that ADF neuron responds reliably to noxious stimuli such as repulsive odors, copper, sodium dodecyl sulfonate (SDS), and mechanical perturbation. This response is mediated by cell-autonomous and non-cell autonomous mechanisms. Furthermore, we show that ADF can modulate avoidance behaviors by inhibiting ASH, an amphid neuron with single ciliated ending. This work greatly furthers our understanding of 5-HT's contributions to sensory information perception, processing, and the resulting behavioral responses.

Bottom-up processing of curvilinear visual features is sufficient for animate/inanimate object categorization.

Animate and inanimate objects differ in their intermediate visual features. For instance, animate objects tend to be more curvilinear compared to inanimate objects (e.g., Levin, Takarae, Miner, & Keil, 2001). Recently, it has been demonstrated that these differences in the intermediate visual features of animate and inanimate objects are sufficient for categorization: Human participants viewing synthesized images of animate and inanimate objects that differ largely in the amount of these visual features classify objects as animate/inanimate significantly above chance (Long, Stormer, & Alvarez, 2017). A remaining question, however, is whether the observed categorization is a consequence of top-down cognitive strategies (e.g., rectangular shapes are less likely to be animals) or a consequence of bottom-up processing of their intermediate visual features, per se, in the absence of top-down cognitive strategies. To address this issue, we repeated the classification experiment of Long et al. (2017) but, unlike Long et al. (2017), matched the synthesized images, on average, in the amount of image-based and perceived curvilinear and rectilinear information. Additionally, in our synthesized images, global shape information was not preserved, and the images appeared as texture patterns. These changes prevented participants from using top-down cognitive strategies to perform the task. During the experiment, participants were presented with these synthesized, texture-like animate and inanimate images and, on each trial, were required to classify them as either animate or inanimate with no feedback given. Participants were told that these synthesized images depicted abstract art patterns. We found that participants still classified the synthesized stimuli significantly above chance even though they were unaware of their classification performance. For both object categories, participants depended more on the curvilinear and less on the rectilinear, image-based information present in the stimuli for classification. Surprisingly, the stimuli most consistently classified as animate were the most dangerous animals in our sample of images. We conclude that bottom-up processing of intermediate features present in the visual input is sufficient for animate/inanimate object categorization and that these features may convey information associated with the affective content of the visual stimuli.

OSM-9 and an amiloride-sensitive channel, but not PKD-2, are involved in mechanosensation in C. elegans male ray neurons.

Mechanotransduction is crucial for touch sensation, hearing, proprioception, and pain sensing. In C. elegans, male ray neurons have been implicated to be involved in the mechanosensation required for mating behavior. However, whether ray neurons directly sense mechanical stimulation is not yet known, and the underlying molecular mechanisms have not been identified. Using in vivo calcium imaging, we recorded the touch-induced calcium responses in male ray neurons. Our data demonstrated that ray neurons are sensitive to mechanical stimulation in a neurotransmitter-independent manner. PKD-2, a putative sensor component for both mechanosensation and chemosensation in male-specific neurons, was not required for the touch-induced calcium responses in RnB neurons, whereas the TRPV channel OSM-9 shaped the kinetics of the responses. We further showed that RnB-neuron mechanosensation is likely mediated by an amiloride-sensitive DEG/ENaC channel. These observations lay a foundation for better understanding the molecular mechanisms of mechanosensation.

TMC Proteins Modulate Egg Laying and Membrane Excitability through a Background Leak Conductance in C. elegans.

Membrane excitability is a fundamentally important feature for all excitable cells including both neurons and muscle cells. However, the background depolarizing conductances in excitable cells, especially in muscle cells, are not well characterized. Although mutations in transmembrane channel-like (TMC) proteins TMC1 and TMC2 cause deafness and vestibular defects in mammals, their precise action modes are elusive. Here, we discover that both TMC-1 and TMC-2 are required for normal egg laying in C. elegans. Mutations in these TMC proteins cause membrane hyperpolarization and disrupt the rhythmic calcium activities in both neurons and muscles involved in egg laying. Mechanistically, TMC proteins enhance membrane depolarization through background leak currents and ectopic expression of both C. elegans and mammalian TMC proteins results in membrane depolarization. Therefore, we have identified an unexpected role of TMC proteins in modulating membrane excitability. Our results may provide mechanistic insights into the functions of TMC proteins in hearing loss and other diseases.

Genome-wide survey of heat shock factors and heat shock protein 70s and their regulatory network under abiotic stresses in Brachypodium distachyon.

The heat shock protein 70s (Hsp70s) and heat shock factors (Hsfs) play key roles in protecting plant cells or tissues from various abiotic stresses. Brachypodium distachyon, recently developed an excellent model organism for functional genomics research, is related to the major cereal grain species. Although B. distachyon genome has been fully sequenced, the information of Hsf and Hsp70 genes and especially the regulatory network between Hsfs and Hsp70s remains incomplete. Here, a total of 24 BdHsfs and 29 BdHsp70s were identified in the genome by bioinformatics analysis and the regulatory network between Hsfs and Hsp70s were performed in this study. Based on highly conserved domain and motif analysis, BdHsfs were grouped into three classes, and BdHsp70s divided into six groups, respectively. Most of Hsf proteins contain five conserved domains: DBD, HR-A/B region, NLS and NES motifs and AHA domain, while Hsp70 proteins have three conserved domains: N-terminal nucleotide binding domain, peptide binding domain and a variable C-terminal lid region. Expression data revealed a large number of BdHsfs and BdHsp70s were induced by HS challenge, and a previous heat acclimation could induce the acquired thermotolerance to help seedling suffer the severe HS challenge, suggesting that the BdHsfs and BdHsp70s played a role in alleviating the damage by HS. The comparison revealed that, most BdHsfs and BdHsp70s genes responded to multiple abiotic stresses in an overlapping relationship, while some of them were stress specific response genes. Moreover, co-expression relationships and predicted protein-protein interaction network implied that class A and B Hsfs played as activator and repressors, respectively, suggesting that BdHsp70s might be regulated by both the activation and the repression mechanisms under stress condition. Our genomics analysis of BdHsfs and BdHsp70s provides important evolutionary and functional characterization for further investigation of the accurate regulatory mechanisms among Hsfs and Hsp70s in herbaceous plants.

Polymodal Responses in C. elegans Phasmid Neurons Rely on Multiple Intracellular and Intercellular Signaling Pathways.

Animals utilize specialized sensory neurons enabling the detection of a wide range of environmental stimuli from the presence of toxic chemicals to that of touch. However, how these neurons discriminate between different kinds of stimuli remains poorly understood. By combining in vivo calcium imaging and molecular genetic manipulation, here we investigate the response patterns and the underlying mechanisms of the C. elegans phasmid neurons PHA/PHB to a variety of sensory stimuli. Our observations demonstrate that PHA/PHB neurons are polymodal sensory neurons which sense harmful chemicals, hyperosmotic solutions and mechanical stimulation. A repulsive concentration of IAA induces calcium elevations in PHA/PHB and both OSM-9 and TAX-4 are essential for IAA-sensing in PHA/PHB. Nevertheless, the PHA/PHB neurons are inhibited by copper and post-synaptically activated by copper removal. Neuropeptide is likely involved in copper removal-induced calcium elevations in PHA/PHB. Furthermore, mechanical stimulation activates PHA/PHB in an OSM-9-dependent manner. Our work demonstrates how PHA/PHB neurons respond to multiple environmental stimuli and lays a foundation for the further understanding of the mechanisms of polymodal signaling, such as nociception, in more complex organisms.

An applet for the Gabor similarity scaling of the differences between complex stimuli.

It is widely accepted that after the first cortical visual area, V1, a series of stages achieves a representation of complex shapes, such as faces and objects, so that they can be understood and recognized. A major challenge for the study of complex shape perception has been the lack of a principled basis for scaling of the physical differences between stimuli so that their similarity can be specified, unconfounded by early-stage differences. Without the specification of such similarities, it is difficult to make sound inferences about the contributions of later stages to neural activity or psychophysical performance. A Web-based app is described that is based on the Malsburg Gabor-jet model (Lades et al., 1993), which allows easy specification of the V1 similarity of pairs of stimuli, no matter how intricate. The model predicts the psycho physical discriminability of metrically varying faces and complex blobs almost perfectly (Yue, Biederman, Mangini, von der Malsburg, & Amir, 2012), and serves as the input stage of a large family of contemporary neurocomputational models of vision.

Curvature-processing network in macaque visual cortex.

Our visual environment abounds with curved features. Thus, the goal of understanding visual processing should include the processing of curved features. Using functional magnetic resonance imaging in behaving monkeys, we demonstrated a network of cortical areas selective for the processing of curved features. This network includes three distinct hierarchically organized regions within the ventral visual pathway: a posterior curvature-biased patch (PCP) located in the near-foveal representation of dorsal V4, a middle curvature-biased patch (MCP) located on the ventral lip of the posterior superior temporal sulcus (STS) in area TEO, and an anterior curvature-biased patch (ACP) located just below the STS in anterior area TE. Our results further indicate that the processing of curvature becomes increasingly complex from PCP to ACP. The proximity of the curvature-processing network to the well-known face-processing network suggests a possible functional link between them.

Neural correlates of personal space intrusion.

A parietal-frontal network in primates is thought to support many behaviors occurring in the space around the body, including interpersonal interactions and maintenance of a particular "comfort zone" or distance from other people ("personal space"). To better understand this network in humans, we used functional MRI to measure the responses to moving objects (faces, cars, simple spheres) and the functional connectivity of two regions in this network, the dorsal intraparietal sulcus (DIPS) and the ventral premotor cortex (PMv). We found that both areas responded more strongly to faces that were moving toward (vs away from) subjects, but did not show this bias in response to comparable motion in control stimuli (cars or spheres). Moreover, these two regions were functionally interconnected. Tests of activity-behavior associations revealed that the strength of DIPS-PMv connectivity was correlated with the preferred distance that subjects chose to stand from an unfamiliar person (personal space size). In addition, the magnitude of DIPS and PMv responses was correlated with the preferred level of social activity. Together, these findings suggest that this parietal-frontal network plays a role in everyday interactions with others.

fMRI analysis of contrast polarity in face-selective cortex in humans and monkeys.

Recognition is strongly impaired when the normal contrast polarity of faces is reversed. For instance, otherwise-familiar faces become very difficult to recognize when viewed as photographic negatives. Here, we used fMRI to demonstrate related properties in visual cortex: 1) fMRI responses in the human Fusiform Face Area (FFA) decreased strongly (26%) to contrast-reversed faces across a wide range of contrast levels (5.3-100% RMS contrast), in all subjects tested. In a whole brain analysis, this contrast polarity bias was largely confined to the Fusiform Face Area (FFA; p<0.0001), with possible involvement of a left occipital face-selective region. 2) It is known that reversing facial contrast affects three image properties in parallel (absorbance, shading, and specular reflection). Here, comparison of FFA responses to those in V1 suggests that the contrast polarity bias is produced in FFA only when all three component properties were reversed simultaneously, which suggests a prominent non-linearity in FFA processing. 3) Across a wide range (180°) of illumination source angles, 3D face shapes without texture produced response constancy in FFA, without a contrast polarity bias. 4) Consistent with psychophysics, analogous fMRI biases for normal contrast polarity were not produced by non-face objects, with image statistics similar to the face stimuli. 5) Using fMRI, we also demonstrated a contrast polarity bias in awake behaving macaque monkeys, in the cortical region considered homologous to human FFA. Thus common cortical mechanisms may underlie facial contrast processing across ~25 million years of primate evolution.

Predicting the psychophysical similarity of faces and non-face complex shapes by image-based measures.

Shape representation is accomplished by a series of cortical stages in which cells in the first stage (V1) have local receptive fields tuned to contrast at a particular scale and orientation, each well modeled as a Gabor filter. In succeeding stages, the representation becomes largely invariant to Gabor coding (Kobatake & Tanaka, 1994). Because of the non-Gabor tuning in these later stages, which must be engaged for a behavioral response (Tong, 2003; Tong et al., 1998), a V1-based measure of shape similarity based on Gabor filtering would not be expected to be highly correlated with human performance when discriminating complex shapes (faces and teeth-like blobs) that differ metrically on a two-choice, match-to-sample task. Here we show that human performance is highly correlated with Gabor-based image measures (Gabor simple and complex cells), with values often in the mid 0.90s, even without discounting the variability in the speed and accuracy of performance not associated with the similarity of the distractors. This high correlation is generally maintained through the stages of HMAX, a model that builds upon the Gabor metric and develops units for complex features and larger receptive fields. This is the first report of the psychophysical similarity of complex shapes being predictable from a biologically motivated, physical measure of similarity. As accurate as these measures were for accounting for metric variation, a simple demonstration showed that all were insensitive to viewpoint invariant (nonaccidental) differences in shape.