Changes In Cellular Excitability Research Articles

Arachidonic acid modulation of α1H, a cloned human T-type calcium channel

Arachidonic acid (AA) and the products of its metabolism are central mediators of changes in cellular excitability. We show that the recently cloned and expressed T-type or low-voltage-activated Ca channel, alpha1H, is modulated by external AA. AA (10 microM) causes a slow, time-dependent attenuation of alpha1H current. At a holding potential of -80 mV, 10 microM AA reduces peak inward alpha1H current by 15% in 15 min and 70% in 30 min and shifts the steady-state inactivation curve -25 mV. AA inhibition was not affected by applying the cyclooxygenase inhibitor indomethacin or the lipoxygenase inhibitor nordihydroguaiaretic acid. The epoxygenase inhibitor octadecynoic acid partially antagonized AA attenuation of alpha1H. The epoxygenase metabolite epoxyeicosatrienoic acid (8,9-EET) mimicked the inhibitory effect of AA on alpha1H peak current. A protein kinase C (PKC)-specific inhibitor (peptide fragment 19-36) only partially antagonized the AA-induced reduction of peak alpha1H current and the shift of the steady-state inactivation curve but had no effect on 8,9-EET-induced attenuation of current. In contrast, PKA has no role in the modulation of alpha1H. These results suggest that AA attenuation and shift of alpha1H may be mediated directly by AA. The heterologous expression of T-type Ca channels allows us to study for the first time properties of this important class of ion channel in isolation. There is a significant overlap of the steady-state activation and inactivation curves, which implies a substantial window current. The selective shift of the steady-state inactivation curve by AA reduces peak Ca current and eliminates the window current. We conclude that AA may partly mediate physiological effects such as vasodilatation via the attenuation of T-type Ca channel current and the elimination of a T-type channel steady window current.

MRNA coding for voltage-gated sodium channel β2 subunit in rat central nervous system: cellular distribution and changes following kainate-induced seizures

The cellular distribution of sodium channel β2 subunit mRNA was examined in the central nervous system from adult Wistar rats using a non-radioactive in situ hybridization method with digoxigenin-labeled cRNA probes. The expression of the subunit was strong in cerebral and cerebellar cortex, in medulla oblongata and in the spinal cord whereas heterogeneous in hippocampus. The distribution was evaluated in hippocampus and cerebral cortex from 1 to 72 h after kainate injection and compared to control rats using densitometric analysis. In these areas, a transient increase was seen 1 h after the drug administration, followed, in the hippocampus, by a significant decrease. These variations differ from those we previously reported for α subunits and might play a role in cellular excitability changes occurring in the course of seizures.

Temporospatial organization of membrane-synaptic modifications and topograms of slow oscillations in cortical potentials during learning

This report concerns the important question of the specificity of the organization of the different stages of formation of a conditioned reflex, considered at the level of membrane-synaptic modifications, which are compared with changes in the spatial organization of slow cortical potentials. A defined sequence of involvement of cellular mechanisms is demonstrated during learning, which may be responsible for various features of the spatial organization of slow cortical potentials. Changes in cellular excitability in cortical neurons are probably connected with oscillations in the overall level of instantaneous topograms and, thus, with the general level of spatial synchronization; the involvement of excitatory and inhibitory synaptic connections (with efficiencies corresponding to different stages in the reflex) may determine the formation of the relief seen in topograms. Parameters at both levels depend on the effects of the hypothalamus on the cortex.

Modeling Hermissenda: I. Differential contributions of I A and I C to type-B cell plasticity

We developed a multicompartmental Hodgkin-Huxley model of the Hermissenda type-B photoreceptor and used it to address the relative contributions of reductions of two K+ currents, IA and IC, to changes in cellular excitability and synaptic strength that occur in these cells after associative learning. We found that reductions of [symbol: see text] C, the peak conductance of IC, substantially increased the firing frequency of the type-B cell during the plateau phase of a simulated light response, whereas reductions of [symbol: see text] A had only a modest contribution to the plateau frequency. This can be understood at least in part by the contributions of these currents to the light-induced (nonspiking) generator potential, the plateau of which was enhanced by [symbol: see text] C reductions, but not by [symbol: see text] A reductions. In contrast, however, reductions of [symbol: see text] A broadened the type-B cell action potential, increased Ca2+ influx, and increased the size of the postsynaptic potential produced in a type-A cell, whereas similar reductions of [symbol: see text] C had only negligible contributions to these measures. These results suggest that reductions of IA and IC play important but different roles in type-B cell plasticity.

Enhancement of type B and A photoreceptor inhibitory synaptic connections in conditioned Hermissenda

Intrinsic changes have been identified in isolated and intact type A and type B photoreceptors following classical conditioning of the nudibranch mollusk Hermissenda. Aspects of various intrinsic, nonsynaptic modifications are expressed by alterations in the excitability of identified photoreceptors in response to the conditioned stimulus. In addition to changes in cellular excitability, changes in synaptic strength between identified neurons have been proposed as a possible mechanism of associative learning in several invertebrate preparations. Here we report that classical conditioning produces differential effects upon the strength of inhibitory monosynaptic connections between identified pairs of type B and type A photoreceptors. The amplitude of IPSPs elicited by an action potential in the medial type B photoreceptor and recorded from medical type A photoreceptors was significantly enhanced in conditioned animals as compared to pseudorandom controls. In contrast, the amplitude of IPSPs elicited by an action potential in the lateral type B photoreceptor and recorded from lateral type A photoreceptors did not show significant synaptic enhancement following conditioning. These results provide additional evidence for differential effects of conditioning upon cellular modifications in identified type A and type B photoreceptors, and further indicate that multiple sites of cellular plasticity exist in the visual system of conditioned Hermissenda.

Cellular mechanisms of delayed recovery of excitability in ventricular tissue

It is well established that ventricular tissue, under some conditions, exhibits the phenomenon of postrepolarization refractoriness (PRR) in which the tissue excitability is depressed after an action potential. We have done parallel experiments on rabbit papillary muscles and on isolated rabbit ventricular cells to explain the cellular basis of this phenomenon, using elevated extracellular K+ concentration ([K+]o) (8 mM) to depolarize the tissue and the isolated cells. For isolated cells, we could separately measure cellular excitability (the inverse of the cellular current threshold) and the cellular responsiveness (the ability of the cell to generate inward current after excitation has occurred). We present two hypotheses that could explain the magnitude and time course of tissue PRR in terms of either changes in cellular excitability or changes in cellular responsiveness. We show that, although small changes in cellular excitability do occur, the predominant cellular mechanism for tissue PRR is the time course of recovery of the cellular responsiveness.

Hormonal regulation of K+-channel messenger RNA in rat myometrium during oestrus cycle and in pregnancy

The dramatic enhancement of smooth-muscle excitability in the uterus which occurs at oestrus and at term in pregnant rats is closely related to increased blood oestrogen concentrations. How oestrogen alters the electrical properties of myometrial cells is unclear, although electrical coupling between cells has been shown to increase. Many examples are known of changes in cellular excitability involving modification of existing ion channels by second-messenger pathways. Steroid hormones, in contrast, are generally thought to influence cellular processes mainly through effects on gene expression, inducing the synthesis of new proteins. Previous work, using an oocyte translation system, has shown that a very slowly activating, voltage-dependent K+ current can be expressed from the poly(A)+ RNA of oestrogen-treated rat uteri. We report here that the messenger RNA species producing this channel is rapidly and reversibly induced in the presence of oestrogen, as shown by the appearance and disappearance of this mRNA during the oestrous cycle, its emergence at the end of pregnancy, and its presence or absence following hormonal treatments. These results suggest that oestrogen controls the expression of a voltage-dependent ion channel in uterine smooth muscle cells.

Increased electrotonic coupling in aged rat hippocampus: A possible mechanism for cellular excitability changes

The effects of aging on the intercellular transfer of the low molecular weight fluorescent dye 5,6-carboxyfluorescein was studied in subfields fascia dentata, CA1, and CA3 of rat hippocampal slices maintained in vitro. All three areas exhibited a statistically significant increase in dye coupling with age. The increased dye coupling was accompanied by an apparent increase in postsynaptic excitability as assessed by the ratio of the population spike to EPSP components of the extracellulary recorded field potential. The possibility that artifactual dye coupling due to cell fusion contributed significantly to these results was ruled out by the demonstrations that a high molecular weight, dextran-coupled fluorescein compound did not produce multiple fills and that dye coupling with carboxyfluorescein could be prevented by prior intracellular loading with Ca++, a procedure that decouples gap junctions in other tissue. The increase in extent of electrical coupling suggested by these data may largely account for the apparent increase in cellular excitability of this tissue with age and may reflect the mechanism by which the senescent hippocampus compensates for the loss of afferent input during the course of normal aging. The additional possibility is raised that increased electrical coupling may reflect a mechanism for permanent associative storage of information in this system.

Classical conditioning rapidly induces specific changes in frequency receptive fields of single neurons in secondary and ventral ectosylvian auditory cortical fields

To determine if learning-induced changes in the response of auditory cortical neurons to a conditioned stimulus (CS) reflect general changes in cellular excitability or alterations in signal processing that are specific to that stimulus, we determined frequency receptive fields (FRFs) of single neurons in secondary and ventral ectosylvian auditory fields of the cat during classical conditioning. Associative changes in FRFs of most cells were specific to the frequency of the CS, established rapidly and reversed by extinction. Thus, learning causes specific changes in cortical processing of sounds whose significance is acquired.

Thalamo-cortical activity during increased gravitational stress

Cortical responses evoked by afferent stimulation of the visual pathway have been observed during increased gravitational stress simulated by a centrifuge. An enhancement of the responses is observed during positive (long axis) acceleration and is related to the magnitude and duration of the linear acceleration. It is not induced by angular accelerations and does not occur during lateral acceleration. It is concluded that the hypotension induced by positive acceleration is essential to the changes in thalamo-cortical excitability. The phenomenon arises from reduced cerebral blood flow but the central effect of carotid receptor activity may be involved. The effects are unlikely to be mediated by the reticular formation but probably arise from changes in cellular excitability induced directly at thalamic and cortical levels. It is suggested that in the conscious animal the increased sensory inflow of gravitational stress may also modify thalamo-cortical excitability, and under these circumstances the effects of increased gravitational stress may modify both reticular and forebrain mechanisms.