A large conductance Ca2+-activated K+ channel in alpha T3-1 pituitary gonadotrophs

The Ca2+-activated K+ channel in endocrine cells is responsible for membrane hyperpolarization and rhythmic firing of action potentials. The probability of opening of this channel is sensitive to intracellular-free Ca2+ concentration. In this study we have identified one such large conductance Ca2+-activated K+ channel in alpha T3-1 pituitary gonadotroph cell. This channel is ohmic with a unit conductance of 170 pS in symmetrical KCl (135 mM) and its current reverses near zero millivolts. When more than one channel is present in the patch membrane they open and close independent of each other, exhibiting no cooperativity between them as expected of a binomial distribution. The regulatory mechanism of this channel in modulating hormone secretion from alpha T3-1 gonadotroph cells is indicated.

Inhibition of Human Two-Pore Domain K+ Channel TREK1 by Local Anesthetic Lidocaine: Negative Cooperativity and Half-of-Sites Saturation Kinetics

TWIK-related K+ channel TREK1, a background leak K+ channel, has been strongly implicated as the target of several general and local anesthetics. Here, using the whole-cell and single-channel patch-clamp technique, we investigated the effect of lidocaine, a local anesthetic, on the human (h) TREK1 channel heterologously expressed in human embryonic kidney 293 cells by an adenoviral-mediated expression system. Lidocaine, at clinical concentrations, produced reversible, concentration-dependent inhibition of hTREK1 current, with IC50 value of 180 mu M, by reducing the single-channel open probability and stabilizing the closed state. We have identified a strategically placed unique aromatic couplet (Tyr352 and Phe355) in the vicinity of the protein kinase A phosphorylation site, Ser348, in the C-terminal domain (CTD) of hTREK1, that is critical for the action of lidocaine. Furthermore, the phosphorylation state of Ser348 was found to have a regulatory role in lidocaine-mediated inhibition of hTREK1. It is interesting that we observed strong intersubunit negative cooperativity (Hill coefficient = 0.49) and half-of-sites saturation binding stoichiometry (half-reaction order) for the binding of lidocaine to hTREK1. Studies with the heterodimer of wild-type (wt)-hTREK1 and Delta 119 C-terminal deletion mutant (hTREK1(wt)-Delta 119) revealed that single CTD of hTREK1 was capable of mediating partial inhibition by lidocaine, but complete inhibition necessitates the cooperative interaction between both the CTDs upon binding of lidocaine. Based on our observations, we propose a model that explains the unique kinetics and provides a plausible paradigm for the inhibitory action of lidocaine on hTREK1.

Transplanting the n-terminus from Kv1.4 to Kv1.1 generates an inwardly rectifying K+ channel

A chimeric channel, 4N/1, was generated from two outwardly rectifying K+ channels by linking the N-terminal cytoplasmic domain of hKv1.4 (N terminus ball and chain of hKv1.4) with the transmembrane body of hKvl.l (Delta 78N1 construct of hKvl.l). The recombinant channel has properties similar to the six transmembrane inward rectifiers and opens on hyperpolarization with a threshold of activation at -90 mV. Outward currents are seen on depolarization provided the channel is first exposed to a hyperpolarizing pulse of -100mV or more. Hyperpolarization at and beyond -130mV provides evidence of channel deactivation. Delta 78N1 does not show inward currents on hyperpolarization but does open on depolarizing from -80mV with characteristics similar to native hKvl.l. The outward currents seen in both Delta 78N1 and 4N/1 inactivate slowly at rates consistent with C-type inactivation. The inward rectification of the 4N/1 chimera is consistent with the inactivation gating mechanism. This implies that the addition of the N-terminus from hKv1.4 to hKvl.l shifts channel activation to hyperpolarizing potentials. These results suggest a mechanism involving the N-terminal cytoplasmic domain for conversion of outward rectifiers to inward rectifiers. (C) 1999 Lippincott Williams & Wilkins.

A naturally occurring amino acid substitution in the voltage-dependent sodium channel selectivity filter affects channel gating

The pore of sodium channels contains a selectivity filter made of 4 amino acids, D/E/K/A. In voltage sensitive sodium channel (Nav) channels from jellyfish to human the fourth amino acid is Ala. This Ala, when mutated to Asp, promotes slow inactivation. In some Nav channels of pufferfishes, the Ala is replaced with Gly. We studied the biophysical properties of an Ala-to-Gly substitution (A1529G) in rat Nav1.4 channel expressed in Xenopus oocytes alone or with a beta 1 subunit. The Ala-to-Gly substitution does not affect monovalent cation selectivity and positively shifts the voltage-dependent inactivation curve, although co-expression with a beta 1 subunit eliminates the difference between A1529G and WT. There is almost no difference in channel fast inactivation, but the beta 1 subunit accelerates WT current inactivation significantly more than it does the A1529G channels. The Ala-to-Gly substitution mainly influences the rate of recovery from slow inactivation. Again, the beta 1 subunit is less effective on speeding recovery of A1529G than the WT. We searched Nav channels in numerous databases and noted at least four other independent Ala-to-Gly substitutions in Nav channels in teleost fishes. Thus, the Ala-to-Gly substitution occurs more frequently than previously realized, possibly under selection for alterations of channel gating.

Quantitative interactions between the A-type K+ current and inositol trisphosphate receptors regulate intraneuronal Ca2+ waves and synaptic plasticity

Key points center dot Active calcium signal propagation occurs when an initial calcium trigger elicits calcium release through endoplasmic reticulum (ER) receptors. A high concentration of the calcium trigger in thin-calibre dendrites would suppress release of calcium through hippocampal inositol trisphosphate receptors (InsP3Rs). center dot Could the high-density expression of A-type K+ channels in thin-calibre dendrites be a mechanism for inhibiting this suppression, thereby restoring the utility of the ER as a substrate for active calcium propagation? center dot Quantitative analyses involving experimentally constrained models reveal a bell-shaped dependence of calcium released through InsP3Rs on the A-type K+ channel density, during the propagation of a calcium wave. center dot A-type K+ channels regulated the relative contribution of ER calcium to the induction of synaptic plasticity in the presence of model metabotropic glutamate receptors. center dot These results identify a novel form of interaction between active dendrites and the ER membrane and suggest that A-type K+ channels are ideally placed for inhibiting the suppression of InsP3Rs in thin-calibre dendrites. Abstract The A-type potassium current has been implicated in the regulation of several physiological processes. Here, we explore a role for the A-type potassium current in regulating the release of calcium through inositol trisphosphate receptors (InsP3R) that reside on the endoplasmic reticulum (ER) of hippocampal pyramidal neurons. To do this, we constructed morphologically realistic, conductance-based models equipped with kinetic schemes that govern several calcium signalling modules and pathways, and constrained the distributions and properties of constitutive components by experimental measurements from these neurons. Employing these models, we establish a bell-shaped dependence of calcium release through InsP3Rs on the density ofA-type potassium channels, during the propagation of an intraneuronal calcium wave initiated through established protocols. Exploring the sensitivities of calcium wave initiation and propagation to several underlying parameters, we found that ER calcium release critically depends on dendritic diameter and that wave initiation occurred at branch points as a consequence of a high surface area to volume ratio of oblique dendrites. Furthermore, analogous to the role ofA-type potassium channels in regulating spike latency, we found that an increase in the density ofA-type potassium channels led to increases in the latency and the temporal spread of a propagating calcium wave. Next, we incorporated kinetic models for the metabotropic glutamate receptor (mGluR) signalling components and a calcium-controlled plasticity rule into our model and demonstrate thatthe presence of mGluRs induced a leftward shift in a BienenstockCooperMunro-like synaptic plasticity profile. Finally, we show that the A-type potassium current could regulate the relative contribution of ER calcium to synaptic plasticity induced either through 900 pulses of various stimulus frequencies or through theta burst stimulation. Our results establish a novel form of interaction between active dendrites and the ER membrane, uncovering a powerful mechanism that could regulate biophysical/biochemical signal integration and steer the spatiotemporal spread of signalling microdomains through changes in dendritic excitability.

Chloroplastic glutamine synthetase from normal and water stressed safflower (Carthamus tinctorius L.) leaves

The chloroplastic isoform of glutamine synthetase (GS(2), EC 6.3.1.2) from normal and water stressed safflower (Carthamus tinctorius L. cv.A-300) leaves has been purified to apparent electrophoretic homogeneity by a procedure involving anion-exchange, hydrophobic and size-exclusion chromatography followed by electroelution of the protein from preparative polyacrylamide gels. The observed molecular weight of the native protein varied from 305-330 kDa depending on the sizing column employed. The native protein is composed of 44 kDa subunits. Under conditions of saturating ammonium and at ATP levels of 0.1-10 mM, double-reciprocal plots with respect to glutamate are biphasic and concave downward at high concentrations of the varied substrate for normal enzyme but are linear for enzyme from water-stressed plants. Under subsaturating ATP levels, K-Glu is over 18-fold lower for enzyme from stressed leaves. The K-m, (ATP) varies with Mg2+ levels in the assay mixture. Double-reciprocal plots of initial velocity with respect to ATP at changing fixed levels of NH4+ are linear for normal enzyme but are curved upwards for enzyme from stressed leaves. Initial velocity data of 1/v vs. 1/ammonium for the enzyme from both the sources are non-linear (curved upwards) when ATP is saturating. At subsaturating ATP levels, the data are linear for normal enzyme but are still non-linear for the enzyme from stressed leaves. The results obtained suggest positively cooperative binding of NH4+ A V-max(/2) value of 3.6 mM for Mg2+ was obtained at 5 mM ATP. The isoelectric point of the native protein from normal and stressed leaves was determined to be, respectively, 5.6 and 6.1. The mixed competitive and competitive inhibitors, methionine sulfoximine and ADP and K-i values of 0.086 mM (0.017 for the enzyme from stressed leaves) and 2.15 mM (1.70 for the enzyme from stressed leaves), respectively. Enzyme from stressed leaves is not inhibited by 5 mM proline. The observed kinetic constants of GS(2) from normal and water stressed safflower seedlings are discussed in relation to the known water-stress tolerance of this crop plant.

Copula-Based Modeling of TMI Brightness Temperature With Rainfall Type

Overland rain retrieval using spaceborne microwave radiometer offers a myriad of complications as land presents itself as a radiometrically warm and highly variable background. Hence, land rainfall algorithms of the Tropical Rainfall Measuring Mission (TRMM) Microwave Imager (TMI) have traditionally incorporated empirical relations of microwave brightness temperature (Tb) with rain rate, rather than relying on physically based radiative transfer modeling of rainfall (as implemented in the TMI ocean algorithm). In this paper, sensitivity analysis is conducted using the Spearman rank correlation coefficient as benchmark, to estimate the best combination of TMI low-frequency channels that are highly sensitive to the near surface rainfall rate from the TRMM Precipitation Radar (PR). Results indicate that the TMI channel combinations not only contain information about rainfall wherein liquid water drops are the dominant hydrometeors but also aid in surface noise reduction over a predominantly vegetative land surface background. Furthermore, the variations of rainfall signature in these channel combinations are not understood properly due to their inherent uncertainties and highly nonlinear relationship with rainfall. Copula theory is a powerful tool to characterize the dependence between complex hydrological variables as well as aid in uncertainty modeling by ensemble generation. Hence, this paper proposes a regional model using Archimedean copulas, to study the dependence of TMI channel combinations with respect to precipitation, over the land regions of Mahanadi basin, India, using version 7 orbital data from the passive and active sensors on board TRMM, namely, TMI and PR. Studies conducted for different rainfall regimes over the study area show the suitability of Clayton and Gumbel copulas for modeling convective and stratiform rainfall types for the majority of the intraseasonal months. Furthermore, large ensembles of TMI Tb (from the most sensitive TMI channel combination) were generated conditional on various quantiles (25th, 50th, 75th, and 95th) of the convective and the stratiform rainfall. Comparatively greater ambiguity was observed to model extreme values of the convective rain type. Finally, the efficiency of the proposed model was tested by comparing the results with traditionally employed linear and quadratic models. Results reveal the superior performance of the proposed copula-based technique.

Interference Alignment Algorithms for the K User Constant MIMO Interference Channel

This paper considers the degrees of freedom (DOF) for a K user multiple-input multiple-output (MIMO) M x N interference channel using interference alignment (IA). A new performance metric for evaluating the efficacy of IA algorithms is proposed, which measures the extent to which the desired signal dimensionality is preserved after zero-forcing the interference at the receiver. Inspired by the metric, two algorithms are proposed for designing the linear precoders and receive filters for IA in the constant MIMO interference channel with a finite number of symbol extensions. The first algorithm uses an eigenbeamforming method to align sub-streams of the interference to reduce the dimensionality of the interference at all the receivers. The second algorithm is iterative, and is based on minimizing the interference leakage power while preserving the dimensionality of the desired signal space at the intended receivers. The improved performance of the algorithms is illustrated by comparing them with existing algorithms for IA using Monte Carlo simulations.

On the generalized degrees of freedom of the K-user symmetric MIMO Gaussian interference channel

This work derives inner and outer bounds on the generalized degrees of freedom (GDOF) of the K-user symmetric MIMO Gaussian interference channel. For the inner bound, an achievable GDOF is derived by employing a combination of treating interference as noise, zero-forcing at the receivers, interference alignment (IA), and extending the Han-Kobayashi (HK) scheme to K users, depending on the number of antennas and the INR/SNR level. An outer bound on the GDOF is derived, using a combination of the notion of cooperation and providing side information to the receivers. Several interesting conclusions are drawn from the bounds. For example, in terms of the achievable GDOF in the weak interference regime, when the number of transmit antennas (M) is equal to the number of receive antennas (N), treating interference as noise performs the same as the HK scheme and is GDOF optimal. For K >; N/M+1, a combination of the HK and IA schemes performs the best among the schemes considered. However, for N/M <; K ≤ N/M+1, the HK scheme is found to be GDOF optimal.

Physical Layer Network Coding for the K-User Multiple Access Relay Channel

We propose a Physical layer Network Coding (PNC) scheme for the K-user wireless Multiple Access Relay Channel, in which K source nodes want to transmit messages to a destination node D with the help of a relay node R. The proposed scheme involves (i) Phase 1 during which the source nodes alone transmit and (ii) Phase 2 during which the source nodes and the relay node transmit. At the end of Phase 1, the relay node decodes the messages of the source nodes and during Phase 2 transmits a many-to-one function of the decoded messages. To counter the error propagation from the relay node, we propose a novel decoder which takes into account the possibility of error events at R. It is shown that if certain parameters are chosen properly and if the network coding map used at R forms a Latin Hypercube, the proposed decoder offers the maximum diversity order of two. Also, it is shown that for a proper choice of the parameters, the proposed decoder admits fast decoding, with the same decoding complexity order as that of the reference scheme based on Complex Field Network Coding (CFNC). Simulation results indicate that the proposed PNC scheme offers a large gain over the CFNC scheme.

On Precoding for Constant K-User MIMO Gaussian Interference Channel With Finite Constellation Inputs

This paper considers linear precoding for the constant channel-coefficient K-user MIMO Gaussian interference channel (MIMO GIC) where each transmitter-i (Tx-i) requires the sending of d(i) independent complex symbols per channel use that take values from fixed finite constellations with uniform distribution to receiver-i (Rx-i) for i = 1, 2, ..., K. We define the maximum rate achieved by Tx-i using any linear precoder as the signal-to-noise ratio (SNR) tends to infinity when the interference channel coefficients are zero to be the constellation constrained saturation capacity (CCSC) for Tx-i. We derive a high-SNR approximation for the rate achieved by Tx-i when interference is treated as noise and this rate is given by the mutual information between Tx-i and Rx-i, denoted as I(X) under bar (i); (Y) under bar (i)]. A set of necessary and sufficient conditions on the precoders under which I(X) under bar (i); (Y) under bar (i)] tends to CCSC for Tx-i is derived. Interestingly, the precoders designed for interference alignment (IA) satisfy these necessary and sufficient conditions. Furthermore, we propose gradient-ascentbased algorithms to optimize the sum rate achieved by precoding with finite constellation inputs and treating interference as noise. A simulation study using the proposed algorithms for a three-user MIMO GIC with two antennas at each node with d(i) = 1 for all i and with BPSK and QPSK inputs shows more than 0.1-b/s/Hz gain in the ergodic sum rate over that yielded by precoders obtained from some known IA algorithms at moderate SNRs.

Inner Bound on the GDOF of the K-User MIMO Gaussian Symmetric Interference Channel

The K-user multiple input multiple output (MIMO) Gaussian symmetric interference channel where each transmitter has M antennas and each receiver has N antennas is studied from a generalized degrees of freedom (GDOF) perspective. An inner bound on the GDOF is derived using a combination of techniques such as treating interference as noise, zero forcing (ZF) at the receivers, interference alignment (IA), and extending the Han-Kobayashi (HK) scheme to K users, as a function of the number of antennas and the log INR/log SNR level. Several interesting conclusions are drawn from the derived bounds. It is shown that when K > N/M + 1, a combination of the HK and IA schemes performs the best among the schemes considered. When N/M < K <= N/M + 1, the HK-scheme outperforms other schemes and is found to be GDOF optimal in many cases. In addition, when the SNR and INR are at the same level, ZF-receiving and the HK-scheme have the same GDOF performance.

Outer Bounds on the Sum Rate of the K-User MIMO Gaussian Interference Channel

This paper derives outer bounds on the sum rate of the K-user MIMO Gaussian interference channel (GIC). Three outer bounds are derived, under different assumptions of cooperation and providing side information to receivers. The novelty in the derivation lies in the careful selection of side information, which results in the cancellation of the negative differential entropy terms containing signal components, leading to a tractable outer bound. The overall outer bound is obtained by taking the minimum of the three outer bounds. The derived bounds are simplified for the MIMO Gaussian symmetric IC to obtain outer bounds on the generalized degrees of freedom (GDOF). The relative performance of the bounds yields insight into the performance limits of multiuser MIMO GICs and the relative merits of different schemes for interference management. These insights are confirmed by establishing the optimality of the bounds in specific cases using an inner bound on the GDOF derived by the authors in a previous work. It is also shown that many of the existing results on the GDOF of the GIC can be obtained as special cases of the bounds, e. g., by setting K = 2 or the number of antennas at each user to 1.

Yielding of sensitive clays: micromechanical considerations

Test results reported on several natural sensitive soils show significant anisotropy of the yield curves, which are generally oriented along the coefficient of earth pressure at rest (K-0) axis. An attempt is made in this paper to explain the anisotropy in yielding from microstructural considerations. An elliptic pore, with particle domains aligned along the periphery of the pore, and with the major axis of the pore being oriented along the direction of the in situ major principal stress, is chosen as the unit of microstructure. An analysis of forces at the interdomain contacts around the ellipse is carried out with reference to experimentally determined yield stress conditions of one soil, and a yield criteria is defined. The analysis, with the proposed yield criteria, enables one to define the complete yield curve for any other soil from the results of only two tests (one constant eta compression test with eta close to eta(K?0), where eta is the stress ratio (= q/p) and eta(K?0) is the stress ratio corresponding to anisotropic K-0 compression, and another undrained shear test). Predicted yield curves are compared with experimental yield curves of several soils reported in the literature.

Einstein Relation in n-Channel Inversion Layers of Nonlinear Optical, Optoelectronic and Related Materials: Simplified Theory, Relative Comparison and Suggestion for an Experimental Determination

In this paper, we study the Einstein relation for the diffusivity to mobility ratio (DMR) in n-channel inversion layers of non-linear optical materials on the basis of a newly formulated electron dispersion relation by considering their special properties within the frame work of k.p formalism. The results for the n-channel inversion layers of III-V, ternary and quaternary materials form a special case of our generalized analysis. The DMR for n-channel inversion layers of II-VI, IV-VI and stressed materials has been investigated by formulating the respective 2D electron dispersion laws. It has been found, taking n-channel inversion layers of CdGeAs2, Cd(3)AS(2), InAs, InSb, Hg1-xCdxTe, In1-xGaxAsyP1-y lattice matched to InP, CdS, PbTe, PbSnTe, Pb1-xSnxSe and stressed InSb as examples, that the DMR increases with the increasing surface electric field with different numerical values and the nature of the variations are totally band structure dependent. The well-known expression of the DMR for wide gap materials has been obtained as a special case under certain limiting conditions and this compatibility is an indirect test for our generalized formalism. Besides, an experimental method of determining the 2D DMR for n-channel inversion layers having arbitrary dispersion laws has been suggested.