The temperature-dependent ion conductance of OmpC, a major outer membrane channel of Escherichia coli, is predicted using all-atom molecular dynamics simulations and experimentally verified. To generalize previous results, OmpC is compared to its structural homolog OmpF at different KCl concentrations, pH values, and a broad temperature range. At low salt concentrations and up to room temperature, the molecular modeling predicts the experimental conductance accurately. At high salt concentrations above 1 M KCl and above room temperature, the simulations underestimate the conductance. Moreover, the temperature dependence of the channel conductance is different from that of the bulk, both in experiment and simulation, indicating a strong contribution of surface effects to the ion conductance. With respect to OmpC, subconductance levels can be observed in experiments only. Subconductance and gating levels can be clearly distinguished by their differences in conductance values and temperature-dependent behavior. With increasing temperature, the probability of a subconductance state to occur, increases, while the dwell time is decreased. The open probability, frequency, and dwell time of such states is largely pH- and KCl concentration-independent, while their amplitudes show a lower increase with increasing salt concentration than gating amplitudes. Voltage dependence of subconductance has been found to be negligible within the uncertainty of the measurements.
In the vertebrate CNS, fast synaptic inhibition is mediated by GABA and glycine receptors. We recently reported that the time course of these synaptic currents is slower when intracellular chloride is high. Here we extend these findings to measure the effects of both extracellular and intracellular chloride on the deactivation of glycine and GABA currents at both negative and positive holding potentials. Currents were elicited by fast agonist application to outside-out patches from HEK-293 cells expressing rat glycine or GABA receptors. The slowing effect of high extracellular chloride on current decay was detectable only in low intracellular chloride (4 mM). Our main finding is that glycine and GABA receptors "sense" chloride concentrations because of interactions between the M2 pore-lining domain and the permeating ions. This hypothesis is supported by the observation that the sensitivity of channel gating to intracellular chloride is abolished if the channel is engineered to become cation selective or if positive charges in the external pore vestibule are eliminated by mutagenesis. The appropriate interaction between permeating ions and channel pore is also necessary to maintain the channel voltage sensitivity of gating, which prolongs current decay at depolarized potentials. Voltage dependence is abolished by the same mutations that suppress the effect of intracellular chloride and also by replacing chloride with another permeant ion, thiocyanate. These observations suggest that permeant chloride affects gating by a foot-in-the-door effect, binding to a channel site with asymmetrical access from the intracellular and extracellular sides of the membrane.
Ensembles of cortical neurons can track fast-varying inputs and relay them in their spike trains, far beyond the cut-off imposed by membrane passive electrical properties and mean firing rates. Initially explored in silico and later demonstrated experimentally, investigating how neurons respond to sinusoidally modulated stimuli provides a deeper insight into spike initiation mechanisms and information processing than conventional F-I curve methodologies. Besides net membrane currents, physiological synaptic inputs can also induce a stimulus-dependent modulation of the total membrane conductance, which is not reproduced by standard current-clamp protocols. Here, we investigated whether rat cortical neurons can track fast temporal modulations over a noisy conductance background. We also determined input-output transfer properties over a range of conditions, including: distinct presynaptic activation rates, postsynaptic firing rates and variability and type of temporal modulations. We found a very broad signal transfer bandwidth across all conditions, similar large cut-off frequencies and power-law attenuations of fast-varying inputs. At slow and intermediate input modulations, the response gain decreased for increasing output mean firing rates. The gain also decreased significantly for increasing intensities of background synaptic activity, thus generalising earlier studies on F-I curves. We also found a direct correlation between the action potentials' onset rapidness and the neuronal bandwidth. Our novel results extend previous investigations of dynamical response properties to non-stationary and conductance-driven conditions, and provide computational neuroscientists with a novel set of observations that models must capture when aiming to replicate cortical cellular excitability.
A new type of lightweight material produced by 3D printing consisting of nano-carbon doped polymer layer followed by a dielectric polymer layer is proposed. We performed temperature dependent characterization and measured the electromagnetic (EM) response of the samples in the GHz and THz range. The temperature dependent structural characteristics, crystallization, and melting were observed to be strongly affected by the presence and the number of nano-carbon doped layers in the sandwich structure. The electromagnetic measurements show a great potential of such a type of periodic material for electromagnetic compatibility applications in microwave frequency range. Sandwich structures containing only two nano-carbon layers already become not transparent to the microwaves, giving an electromagnetic interference shielding efficiency at the level of 8-15 dB. A sandwich consisting of one nano-carbon doped and one polymer layer is opaque for THz radiation, because of 80% of absorption. These studies serve as a basis for design and realization of specific optimal geometries of meta-surface type with the 3D printing technique, in order to reach a high level of electromagnetic interference shielding performance for real world EM cloaking and EM ecology applications. V
Firing rate variability at the single neuron level is characterized by long-memory processes and complex statistics over a wide range of time scales (from milliseconds up to several hours). Here, we focus on the contribution of non-stationary efficacy of the ensemble of synapses–activated in response to a given stimulus–on single neuron response variability. We present and validate a method tailored for controlled and specific long-term activation of a single cortical neuron in vitro via synaptic or antidromic stimulation, enabling a clear separation between two determinants of neuronal response variability: membrane excitability dynamics vs. synaptic dynamics. Applying this method we show that, within the range of physiological activation frequencies, the synaptic ensemble of a given neuron is a key contributor to the neuronal response variability, long-memory processes and complex statistics observed over extended time scales. Synaptic transmission dynamics impact on response variability in stimulation rates that are substantially lower compared to stimulation rates that drive excitability resources to fluctuate. Implications to network embedded neurons are discussed.
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