Channel Sensing Research Articles

Marine High-Speed Over-the-Horizon Communications and Channel Sensing in Evaporation Ducts Over the South China Sea

Marine High-Speed Over-the-Horizon Communications and Channel Sensing in Evaporation Ducts Over the South China Sea

Scorpion α-toxin LqhαIT specifically interacts with a glycan at the pore domain of voltage-gated sodium channels

Voltage-gated sodium (Nav) channels sense membrane potential and drive cellular electrical activity. The deathstalker scorpion α-toxin LqhαIT exerts a strong action potential prolonging effect on Nav channels. To elucidate the mechanism of action of LqhαIT, we determined a 3.9Å cryoelectron microscopy (cryo-EM) structure of LqhαIT in complex with the Nav channel from Periplaneta americana (NavPas). We found that LqhαIT binds to voltage sensor domain 4 and traps it in an "S4 down" conformation. The functionally essential C-terminal epitope of LqhαIT forms an extensive interface with the glycan scaffold linked to Asn330 of NavPas that augments a small protein-protein interface between NavPas and LqhαIT. A combination of molecular dynamics simulations, structural comparisons, and prior mutagenesis experiments demonstrates the functional importance of this toxin-glycan interaction. These findings establish a structural basis for the specificity achieved by scorpion α-toxins and reveal the conserved glycan as an essential component of the toxin-binding epitope.

Fine-tuning pH sensor H98 by remote essential residues in the hydrogen-bond network of mTASK-3

TASK-3 generates a background K+ conductance which when inhibited by acidification depolarizes membrane potential and increases cell excitability. These channels sense pH by protonation of histidine residue H98, but recent evidence revealed that several other amino acid residues also contribute to TASK-3 pH sensitivity, suggesting that the pH sensitivity is determined by an intermolecular network. Here we use electrophysiology and molecular modeling to characterize the nature and requisite role(s) of multiple amino acids in pH sensing by TASK-3. Our results suggest that the pH sensor H98 and consequently pH sensitivity is influenced by remote amino acids that function as a hydrogen-bonding network to modulate ionic conductivity. Among the residues in the network, E30 and K79 are the most important for passing external signals near residue S31 to H98. The hydrogen-bond network plays a key role in selectivity or pH sensing in mTASK-3, and E30 and S31 in the network can modulate the conductive properties (E30) or reverse the pH sensitivity and selectivity of the channel (S31). Molecular dynamics simulations and pK1/2 calculation revealed that double mutants involving H98 + S31 primarily regulate the structure stability of the pore selectivity filter and pore loop regions, further strengthen the stability of the cradle suspension system, and alter the ionization state of E30 and K79, thereby preventing pore conformational change that normally occurs in response to varying extracellular pH. These results demonstrate that crucial residues in the hydrogen-bond network can remotely tune the pH sensing of mTASK-3 and may be a potential allosteric regulatory site for therapeutic molecule development.

Piezo1 channels restrain ILC2s and regulate the development of airway hyperreactivity.

Mechanosensitive ion channels sense force and pressure in immune cells to drive the inflammatory response in highly mechanical organs. Here, we report that Piezo1 channels repress group 2 innate lymphoid cell (ILC2)-driven type 2 inflammation in the lungs. Piezo1 is induced on lung ILC2s upon activation, as genetic ablation of Piezo1 in ILC2s increases their function and exacerbates the development of airway hyperreactivity (AHR). Conversely, Piezo1 agonist Yoda1 reduces ILC2-driven lung inflammation. Mechanistically, Yoda1 inhibits ILC2 cytokine secretion and proliferation in a KLF2-dependent manner, as we found that Piezo1 engagement reduces ILC2 oxidative metabolism. Consequently, in vivo Yoda1 treatment reduces the development of AHR in experimental models of ILC2-driven allergic asthma. Human-circulating ILC2s express and induce Piezo1 upon activation, as Yoda1 treatment of humanized mice reduces human ILC2-driven AHR. Our studies define Piezo1 as a critical regulator of ILC2s, and we propose the potential of Piezo1 activation as a novel therapeutic approach for the treatment of ILC2-driven allergic asthma.

Loss of the auxiliary α2δ1 voltage-sensitive calcium channel subunit impairs bone formation and anabolic responses to mechanical loading.

Voltage-sensitive calcium channels (VSCCs) influence bone structure and function, including anabolic responses to mechanical loading. While the pore-forming (α1) subunit of VSCCs allows Ca2+ influx, auxiliary subunits regulate the biophysical properties of the pore. The α2δ1 subunit influences gating kinetics of the α1 pore and enables mechanically induced signaling in osteocytes; however, the skeletal function of α2δ1 in vivo remains unknown. In this work, we examined the skeletal consequences of deleting Cacna2d1, the gene encoding α2δ1. Dual-energy X-ray absorptiometry and microcomputed tomography imaging demonstrated that deletion of α2δ1 diminished bone mineral content and density in both male and female C57BL/6 mice. Structural differences manifested in both trabecular and cortical bone for males, while the absence of α2δ1 affected only cortical bone in female mice. Deletion of α2δ1 impaired skeletal mechanical properties in both sexes, as measured by three-point bending to failure. While no changes in osteoblast number or activity were found for either sex, male mice displayed a significant increase in osteoclast number, accompanied by increased eroded bone surface and upregulation of genes that regulate osteoclast differentiation. Deletion of α2δ1 also rendered the skeleton insensitive to exogenous mechanical loading in males. While previous work demonstrates that VSCCs are essential for anabolic responses to mechanical loading, the mechanism by which these channels sense and respond to force remained unclear. Our data demonstrate that the α2δ1 auxiliary VSCC subunit functions to maintain baseline bone mass and strength through regulation of osteoclast activity and also provides skeletal mechanotransduction in male mice. These data reveal a molecular player in our understanding of the mechanisms by which VSCCs influence skeletal adaptation.

Channel Sensing for Holographic Interference Surfaces based on the Principle of Interferometry

Channel Sensing for Holographic Interference Surfaces based on the Principle of Interferometry

The role of SIC on the design of next generation multiple access

The interplay of physical layer enhancements and classic random access protocols is the objective of this paper. Successive interference cancellation (SIC) is among the major enhancements of the physical layer. Considering the classic representatives of random access protocols, Slotted ALOHA and Channel Sensing Multiple Access (CSMA), we show that two regimes can be identified as a function of the communication link spectral efficiency. In case of high levels of spectral efficiency, multi-packet reception enabled by SIC is of limited benefit. Sum-rate performance is dominated by the effectiveness of the Medium Access Control (MAC) protocol. On the contrary, for low spectral efficiency levels, sum-rate performance is essentially dependent on physical layer SIC capability, while the MAC protocol has a marginal impact. Limitations due to transmission power dynamic range are shown to induce unfairness among nodes. However, the unfairness issue fades away when the system is driven to work around the sum-rate peak achieved for low spectral efficiency. This can also be confirmed by looking at Age of Information (AoI) metric. The major finding of this work is that SIC can boost performance, while still maintaining a fair sharing of the communication channel among nodes. In this regime, the MAC protocol appears to play a marginal role, while multi-packet reception endowed by SIC is prominent to provide high sum-rate, low energy consumption, and low AoI.

Approaches for the modulation of mechanosensitive MscL channel pores.

MscL was the first mechanosensitive ion channel identified in bacteria. The channel opens its large pore when the turgor pressure of the cytoplasm increases close to the lytic limit of the cellular membrane. Despite their ubiquity across organisms, their importance in biological processes, and the likelihood that they are one of the oldest mechanisms of sensory activation in cells, the exact molecular mechanism by which these channels sense changes in lateral tension is not fully understood. Modulation of the channel has been key to understanding important aspects of the structure and function of MscL, but a lack of molecular triggers of these channels hindered early developments in the field. Initial attempts to activate mechanosensitive channels and stabilize functionally relevant expanded or open states relied on mutations and associated post-translational modifications that were often cysteine reactive. These sulfhydryl reagents positioned at key residues have allowed the engineering of MscL channels for biotechnological purposes. Other studies have modulated MscL by altering membrane properties, such as lipid composition and physical properties. More recently, a variety of structurally distinct agonists have been shown bind to MscL directly, close to a transmembrane pocket that has been shown to have an important role in channel mechanical gating. These agonists have the potential to be developed further into antimicrobial therapies that target MscL, by considering the structural landscape and properties of these pockets.

The roles of two extracellular loops in proton sensing and permeation in human Otop1 channel.

Otopetrin (Otop) proteins were recently found to function as proton channels, with Otop1 revealed to be the sour taste receptor in mammals. Otop proteins contain twelve transmembrane segments (S1-S12) which are divided into structurally similar N and C domains. The mechanisms by which Otop channels sense extracellular protons to initiate gating and conduct protons once the channels are activated remains largely elusive. Here we show that two extracellular loops are playing key roles in human Otop1 channel function. We find that the S5-S6 loop is critical for proton sensing of Otop1. Further, our data reveal that the S11-12 loop is structurally and functionally essential for the Otop1 channel and this loop regulates proton permeation into the pore formed by the C domain. This study sheds light on the molecular mechanism behind the structure and function of this newly identified ion channel family.

How does fast-inactivation PIEZO1 channel sense constant mechanical forces in native endothelial cells?

Mechanical forces including shear stress and hydrostatic pressure play pivotal roles in endothelial physiology and pathophysiology. Native endothelial cells are constantly exposed to these mechanical forces, while most mechanosensitive ion channels including the notable PIEZO1 channels possess a well-known fast-inactivation kinetic. It remains completely unknown how the constant mechanical forces generated by shear stress or hydrostatic pressure are sensed in endothelium. In this study, we identified that PIEZO1 channels are the mechanosensors for both shear stress and hydrostatic pressure in native endothelial cells through pharmacological and gene manipulation approaches. Interestingly, these channels have an unusual non-inactivation property. We further revealed the biophysical basis underlying this conversion of fast-inactivation to non-inactivation in endothelial PIEZO1 channel through electrophysiology, computational modelling and an unbiased screening. It was found that inhibition of neutral sphingomyelinase activity cause PIEZO1 to switch to profoundly inactivating behaviour in native endothelial cells. Moreover, a key product of neutral sphingomyelinase, ceramide, rescues non-inactivating channel behaviour, while its co-product, phosphoryl choline, had no effect. In contrast to ceramide, the substrate of neutral sphingomyelinase, sphingomyelin, did not affect inactivation but altered the channel's mechanosensitivity. Molecular dynamics simulations suggested that the reduction of the depth of the inward-facing dome in endothelial cell membrane caused by sphingomyelinase activity through replacing sphingomyelin with newly-generated ceramide could underlie this atypical transition of PIEZO1 inactivation kinetic. The data suggest that local lipid environment created by sphingomyelinase activity and ceramide enables fast-inactivation PIEZO1 channel to sense constant mechanical forces in native endothelial cells through a novel biophysical and regulatory mechanism.

Charge-voltage curves of Shaker potassium channel are not hysteretic at steady state.

Charge-voltage curves of many voltage-gated ion channels exhibit hysteresis but such curves are also a direct measure of free energy of channel gating and, hence, should be path-independent. Here, we identify conditions to measure steady-state charge-voltage curves and show that these are curves are not hysteretic. Charged residues in transmembrane segments of voltage-gated ion channels (VGICs) sense and respond to changes in the electric field. The movement of these gating charges underpins voltage-dependent activation and is also a direct metric of the net free-energy of channel activation. However, for most voltage-gated ion channels, the charge-voltage (Q-V) curves appear to be dependent on initial conditions. For instance, Q-V curves of Shaker potassium channel obtained by hyperpolarizing from 0 mV is left-shifted compared to those obtained by depolarizing from a holding potential of -80 mV. This hysteresis in Q-V curves is a common feature of channels in the VGIC superfamily and raises profound questions about channel energetics because the net free-energy of channel gating is a state function and should be path independent. Due to technical limitations, conventional gating current protocols are limited to test pulse durations of <500 ms, which raises the possibility that the dependence of Q-V on initial conditions reflects a lack of equilibration. Others have suggested that the hysteresis is fundamental thermodynamic property of voltage-gated ion channels and reflects energy dissipation due to measurements under non-equilibrium conditions inherent to rapid voltage jumps (Villalba-Galea. 2017. Channels. https://doi.org/10.1080/19336950.2016.1243190). Using an improved gating current and voltage-clamp fluorometry protocols, we show that the gating hysteresis arising from different initial conditions in Shaker potassium channel is eliminated with ultra-long (18-25 s) test pulses. Our study identifies a modified gating current recording protocol to obtain steady-state Q-V curves of a voltage-gated ion channel. Above all, these findings demonstrate that the gating hysteresis in Shaker channel is a kinetic phenomenon rather than a true thermodynamic property of the channel and the charge-voltage curve is a true measure of the net-free energy of channel gating.

Controllable Wireless Spoofing Attack Based on Conditional BEGAN and Auxiliary Channel Sensing

This paper investigates how to build a controllable wireless spoofing attack launch framework that is driven by fundamental channel modeling and practical wireless datasets. First, we propose a wireless spoofing attack scheme against the defense mechanism with adversarial deep learning. To obtain channel characteristics and facilitate offline training of the attack model, auxiliary channel sensing is proposed with fundamental channel modeling. Based on these, a conditional boundary equilibrium generative adversarial network (CBEGAN) is designed with adversarial autoencoder (AAE), which takes true labels of signals and channel characteristics as conditions and enables the generation of controllable spoofing signals to fool the protected legitimate classifier. We verify the performance of the proposed spoofing attack scheme with CBEGAN and channel sensing by using wireless datasets, which contain signal data of multiple emitters and modulation types. Results show that the proposed scheme outperforms random attack, replay attack, and the recent attack scheme based on generative adversarial network (GAN) when a single legitimate emitter sends a fixed modulation type. It is also shown that the average attack success probability of the proposed CBEGAN attack model can reach more than 80% while mimicking multiple emitters and modulation types. The performance of the proposed scheme on different channel conditions including signal-to-noise ratio (SNR) and K-factor of the Rician fading channel is evaluated.

Deep Learning for Channel Sensing and Hybrid Precoding in TDD Massive MIMO OFDM Systems

This paper proposes a deep learning approach to channel sensing and downlink hybrid beamforming for massive multiple-input multiple-output systems operating in the time division duplex mode and employing either single-carrier or multicarrier transmission. The conventional precoding design involves a two-step process of first estimating the high-dimensional channel, then designing the precoders based on such estimate. This two-step process is, however, not necessarily optimal. This paper shows that by using a learning approach to design the analog sensing and the hybrid downlink precoders directly from the received pilots without the intermediate high-dimensional channel estimation, the overall system performance can be significantly improved. Training a neural network to design the analog and digital precoders simultaneously is, however, difficult. Further, such an approach is not generalizable to systems with different number of users. In this paper, we develop a simplified and generalizable approach that learns the uplink sensing matrix and downlink analog precoder using a deep neural network that decomposes on a per-user basis, then designs the digital precoder based on the estimated low-dimensional equivalent channel. Numerical comparisons show that the proposed methodology results in significantly less training overhead and leads to an architecture that generalizes to various system settings.

The roles of two extracellular loops in proton sensing and permeation in human Otop1 proton channel

Otopetrin (Otop) proteins were recently found to function as proton channels, with Otop1 revealed to be the sour taste receptor in mammals. Otop proteins contain twelve transmembrane segments (S1-S12) which are divided into structurally similar N and C domains. The mechanisms by which Otop channels sense extracellular protons to initiate gating and conduct protons once the channels are activated remains largely elusive. Here we show that two extracellular loops are playing key roles in human Otop1 channel function. We find that residue H229 in the S5-S6 loop is critical for proton sensing of Otop1. Further, our data reveal that the S11-12 loop is structurally and functionally essential for the Otop1 channel and that residue D570 in this loop regulates proton permeation into the pore formed by the C domain. This study sheds light on the molecular mechanism behind the structure and function of this newly identified ion channel family.

The human TRPA1 intrinsic cold and heat sensitivity involves separate channel structures beyond the N-ARD domain

TRP channels sense temperatures ranging from noxious cold to noxious heat. Whether specialized TRP thermosensor modules exist and how they control channel pore gating is unknown. We studied purified human TRPA1 (hTRPA1) truncated proteins to gain insight into the temperature gating of hTRPA1. In patch-clamp bilayer recordings, ∆1–688 hTRPA1, without the N-terminal ankyrin repeat domain (N-ARD), was more sensitive to cold and heat, whereas ∆1–854 hTRPA1, also lacking the S1–S4 voltage sensing-like domain (VSLD), gained sensitivity to cold but lost its heat sensitivity. In hTRPA1 intrinsic tryptophan fluorescence studies, cold and heat evoked rearrangement of VSLD and the C-terminus domain distal to the transmembrane pore domain S5–S6 (CTD). In whole-cell electrophysiology experiments, replacement of the CTD located cysteines 1021 and 1025 with alanine modulated hTRPA1 cold responses. It is proposed that hTRPA1 CTD harbors cold and heat sensitive domains allosterically coupled to the S5–S6 pore region and the VSLD, respectively.

3D Printed Platform for Impedimetric Sensing of Liquids and Microfluidic Channels.

Fused deposition modeling 3D printing (FDM-3DP) employing electrically conductive filaments has recently been recognized as an exceptionally attractive tool for the manufacture of sensing devices. However, capabilities of 3DP electrodes to measure electric properties of materials have not yet been explored. To bridge this gap, we employ bimaterial FDM-3DP combining electrically conductive and insulating filaments to build an integrated platform for sensing conductivity and permittivity of liquids by impedance measurements. The functionality of the device is demonstrated by measuring conductivity of aqueous potassium chloride solution and bottled water samples and permittivity of water, ethanol, and their mixtures. We further implement an original idea of applying impedance measurements to investigate dimensions of 3DP channels as base structures of microfluidic devices, complemented by their optical microscopic analysis. We demonstrate that FDM-3DP allows the manufacture of microchannels of width down to 80 μm.

Beamspace ESPRIT for mmWave Channel Sensing: Performance Analysis and Beamformer Design

In this paper, we consider the beamspace ESPRIT algorithm for Millimeter-Wave (mmWave) channel sensing. We provide a non-asymptotic analysis of the beamspace ESPRIT algorithm. We derive a deterministic upper bound for the matching distance error between the true angle of arrival (AoA) of the channel paths and the estimated AoA considering a bounded noise model. Additionally, we leverage the insight obtained from our theoretical analysis to propose a novel max-min criterion for beamformer design which can enhance the performance of mmWave channel estimation algorithms, including beamspace ESPRIT. We consider a family of multi-resolution beamformers which can be implemented using phase shifters and introduce a design scheme for the optimal beamformers from this family with respect to the proposed max-min criteria. We can guarantee a minimum beamforming gain uniformly over a region of possible multipath directions, which can lead to more robust channel estimation. We provide several numerical experiments to verify our theoretical claims and demonstrate the superior performance of the proposed beamformers compared to existing beamformer design criteria.

TRP channels: Intestinal bloating TRiPs up pathogen avoidance

Transient receptor potential (TRP) channels are expressed in many nonneural tissues where their functions are not well known. Using C. elegans as a model, a new study demonstrated that colonization of the Gram-positive pathogenic bacteria E. faecalis in the intestine causes intestinal distention. Two TRPM channels sense such intestinal distension to trigger fast pathogen avoidance behavior, thereby limiting pathogen infection. This work signifies the novel role of TRP channels in gut physiology and pathogen defense.

Visualization of the mechanosensitive ion channel MscS under membrane tension.

Mechanosensitive channels sense mechanical forces in cell membranes and underlie many biological sensing processes1-3. However, how exactly they sense mechanical force remains under investigation4. The bacterial mechanosensitive channel of small conductance, MscS, is one of the most extensively studied mechanosensitive channels4-8, but how it is regulated by membrane tension remains unclear, even though the structures are known for its open and closed states9-11. Here we used cryo-electron microscopy to determine the structure of MscS in different membrane environments, including one that mimics a membrane under tension. We present the structures of MscS in the subconducting and desensitized states, and demonstrate that the conformation of MscS in a lipid bilayer in the open state is dynamic. Several associated lipids have distinct roles in MscS mechanosensation. Pore lipids are necessary to prevent ion conduction in the closed state. Gatekeeper lipids stabilize the closed conformation and dissociate with membrane tension, allowing the channel to open. Pocket lipids in a solvent-exposed pocket between subunits are pulled out under sustained tension, allowing the channel to transition to the subconducting state and then to the desensitized state. Our results provide a mechanistic underpinning and expand on the 'force-from-lipids' model for MscS mechanosensation4,11.

Small-conductance Ca2+-activated K+ channels promote J-wave syndrome and phase 2 reentry

Small-conductance Ca2+-activated potassium (SK) channels play complex roles in cardiac arrhythmogenesis. SK channels colocalize with L-type Ca2+ channels, yet how this colocalization affects cardiac arrhythmogenesis is unknown. The purpose of this study was to investigate the role of colocalization of SK channels with L-type Ca2+ channels in promoting J-wave syndrome and ventricular arrhythmias. We carried out computer simulations of single-cell and tissue models. SK channels in the model were assigned to preferentially sense Ca2+ in the bulk cytosol, subsarcolemmal space, or junctional cleft. When SK channels sense Ca2+ in the bulk cytosol, the SK current (ISK) rises and decays slowly during an action potential, the action potential duration (APD) decreases as the maximum conductance increases, no complex APD dynamics and phase 2 reentry canbe induced by ISK. When SK channels sense Ca2+ in the subsarcolemmal space or junctional cleft, ISK can rise and decay rapidly during an action potential in a spike-like pattern because of spiky Ca2+ transients in these compartments, which can cause spike-and-dome action potential morphology, APD alternans, J-wave elevation, and phase 2 reentry. Our results can account for the experimental finding that activation of ISK induced J-wave syndrome and phase 2 reentry in rabbit hearts. Colocalization of SK channels with L-type Ca2+ channels so that they preferentially sense Ca2+ in the subsarcolemmal or junctional space may result in a spiky ISK, which can functionally play a similar role of the transient outward K+ current in promoting J-wave syndrome and ventricular arrhythmias.