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J R Clay

Publications and source records attributed to J R Clay.

At least 19 recordsLinked to original sources
PubMed2002

Temperature dependence of bistability in squid giant axons with alkaline intracellular pH.

Raising the intracellular pH (pHi) above 7.7 in intracellularly perfused squid giant axons causes spontaneous firing of action potentials. The firing frequency ranged from 20 Hz at 0 degrees C to 200 Hz at 23 degrees C. Above 23 degrees C, the axons were quiescent. They were bistable for 13 <T <23 degrees C. That is, they were either quiescent or spontaneously firing. Below 13 degrees C, spontaneous firing was the only stable element. The primary effects of changes in temperature on the underlying ionic currents were on gating of the delayed rectifier potassium channel IK, and the sodium ion channel INa. The kinetics of IK had a Q10 of 3.63. The effect of T on INa was more complicated in that the peak INa amplitude increased with T, as demonstrated in earlier reports. This effect, as well as the changes in INa kinetics produced by changes in T, were mimicked in the context of a model of INa gating in which activation and inactivations are coupled. Electrical activity was simulated in a model of the action potential with appropriate temperature-dependent modifications for INa and IK. The model predicts a change from monostability (spontaneous firing) at relatively low temperatures to bistability (quiescence and spontaneous firing) as the temperature is raised, followed by change back to monostability (quiescence) as the temperature is further increased, which is consistent with the experimental results.

Action Potentials↗
PubMed2001

Action potentials occur spontaneously in squid giant axons with moderately alkaline intracellular pH.

This report demonstrates a novel finding from the classic giant axon preparation of the squid. Namely, the axon can be made to fire autonomously (spontaneously occurring action potentials) when the intracellular pH (pH(i)) was increased to about 7.7, or higher. (Physiological pH(i) is 7.3.) The frequency of firing was 33 Hz (T = 5 degrees ). No changes in frequency or in the voltage waveform itself were observed when pH(i) was increased from 7.7 up to 8.5. In other words, the effect has a threshold at a pH(i) of about 7.7. A mathematical model that is sufficient to mimic these results is provided using a modified version of the Clay (1998) description of the axonal ionic currents.

Action Potentials↗
PubMed2000

Localization of voltage-gated K(+) channels in squid giant axons.

We have localized the classical voltage-gated K(+) channel within squid giant axons by immunocytochemistry using the Kv1 antibody of Rosenthal et al. (1996). Widely dispersed patches of intense immunofluorescence were observed in the axonal membrane. Punctate immunofluorescence was also observed in the axoplasm and was localized to approximately 25-50-microm-wide column down the length of the nerve (axon diameter approximately 500 microm). Immunoelectronmicroscopy of the axoplasm revealed a K(+) channel containing vesicles, 30-50 nm in diameter, within this column. These and other vesicles of similar size were isolated from axoplasm using a novel combination of high-speed ultracentrifugation and controlled-pore size, glass bead separation column techniques. Approximately 1% of all isolated vesicles were labeled by K(+) channel immunogold reacted antibody. Incorporation of isolated vesicle fractions within an artificial lipid bilayer revealed K(+) channel electrical activity similar to that recorded directly from the axonal membrane by Llano et al. (1988). These K(+) channel-containing vesicles may be involved in cycling of K(+) channel protein into the axonal membrane. We have also isolated an axoplasmic fraction containing approximately 150-nm-diameter vesicles that may transport K(+) channels back to the cell body.

Animals↗
PubMed2000

Determining K+ channel activation curves from K+ channel currents.

Potassium ion channels are generally believed to have current-voltage (IV) relations which are linearly related to driving force ( V - E(K)), where V is membrane potential and E(K) is the potassium ion equilibrium potential. Consequently, activation curves for K+ channels have often been measured by normalizing voltage-clamp families of macroscopic K+ currents with (V - E(K)), where V is the potential of each successive step in the voltage clamp sequence. However, the IV relation for many types of K+ channels actually has a non-linear dependence upon driving force which is well described by the Goldman-Hodgkin-Katz relation. When the GHK dependence on (V - E(K)) is used in the normalization procedure, a very different voltage dependence of the activation curve is obtained which may more accurately reflect this feature of channel gating. Novel insights into the voltage dependence of the rapidly inactivating I(A) channels Kv1.4 and Kv4.2 have been obtained when this procedure was applied to recently published results.

Animals↗
PubMed1999

On the role of subthreshold dynamics in neuronal signaling.

The role of subthreshold dynamics in neuronal signaling is examined using periodic pulse train stimulation of the Fitzhugh-Nagumo (FN) model of nerve membrane excitability and results from the squid giant axon as an experimental data base. For a broad range of stimulus conditions the first pulse in a pulse train elicited an action potential, whereas all subsequent pulses elicited subthreshold responses, both in the axon and in the FN model. These results are not well described by the Hodgkin and Huxley 1952 model. Various different patterns of subthreshold responses, including chaotic dynamics, can be observed in both systems-the FN model and the axon-depending upon stimulus conditions. For some conditions action potentials are occasionally interspersed among the subthreshold events with randomly occurring interspike intervals. The randomness is directly attributable to the underlying subthreshold chaos-deterministic chaos-rather than to a stochastic noise source. We conclude that this mechanism may contribute to multimodal interspike interval histograms which have been observed from individual neurons throughout the nervous system.

Action Potentials↗
PubMed1999

Loss of IA expression and increased excitability in postnatal rat Cajal-Retzius cells.

Although an important secretory function of Cajal-Retzius (CR) cells has been discovered recently, the precise electrical status of these cells among other layer I neurons in particular and in cortical function in general is still unclear. In this paper, early postnatal CR cells from rat neocortex were found to express an inactivating K current whose molecular substrate is likely to be the Kv1.4 channel. Both electrophysiological and immunocytochemical experiments revealed that expression of this A-type current is down-regulated in vivo and virtually disappears by the end of the second postnatal week. At this time, CR cells have become capable of evoked repetitive firing, and their action potentials are larger and faster, yet these electrical properties still appear incompatible with a role in cortical network function, as inferred from comparisons with other cortical neurons. Also at this time, a large proportion of CR cells display spontaneous spiking activity, which suggests the possibility of additional roles for these cells. We conclude that the loss of A channels along with an increase in Na channel density shape the changes in excitability of postnatal CR cells, in terms of both the patterns of evoked firing and the emergence of spontaneous spiking.

Action Potentials↗
PubMed1998

Prenatal expression of inwardly rectifying potassium channel mRNA (Kir4.1) in rat brain.

The levels and cellular localization of the mRNA encoding the inwardly rectifying potassium ion channel Kir4.1 were investigated in the embryonic rat brain by Northern blots and in situ hybridization. This transcript was absent at embryonic day 13 (E13), whereas it was clearly present in E14-15 preparations, principally in the neuroepithelium of the cerebral cortex, thalamus, and hypothalamus. At later embryonic stages (E17-20), Kir4.1 mRNA levels increased and expanded to the mantle zone, such as the cortical plate, hippocampus, thalamus, and hypothalamus. The early appearance of Kir4.1 mRNA in various brain regions suggests an involvement of the channel in cell proliferation, migration and differentiation in the rat CNS.

Animals↗
PubMed1998

Effects of divalent cations on the E-4031-sensitive repolarization current, I(Kr), in rabbit ventricular myocytes.

The effects of divalent cations on the E-4031-sensitive repolarization current (I(Kr)) were studied in single ventricular myocytes isolated from rabbit hearts. One group of divalent cations (Cd2+, Ni2+, Co2+, and Mn2+) produced a rightward shift of the I(Kr) activation curve along the voltage axis, increased the maximum I(Kr) amplitude (i.e., relieved the apparent inward rectification of the channel), and accelerated I(Kr) tail current kinetics. Another group (Ca2+, Mg2+ and Sr2+) had relatively little effect on I(Kr). The only divalent cation that blocked I(Kr) was Zn2+ (0.1-1 mM). Under steady-state conditions, Ba2+ caused a substantial block of I(K1) as previously reported. However, block by Ba2+ was time dependent, which precluded a study of Ba2+ effects on I(Kr). We conclude that the various effects of the divalent cations can be attributed to interactions with distinct sites associated with the rectification and/or inactivation mechanism of the channel.

Animals↗
PubMed1998

Excitability of the squid giant axon revisited.

The electrical properties of the giant axon from the common squid Loligo pealei have been reexamined. The primary motivation for this work was the observation that the refractoriness of the axon was significantly greater than the predictions of the standard model of nerve excitability. In particular, the axon fired only once in response to a sustained, suprathreshold stimulus. Similarly, only a single action potential was observed in response to the first pulse of a train of 1-ms duration current pulses, when the pulses were separated in time by approximately 10 ms. The axon was refractory to all subsequent pulses in the train. The underlying mechanisms for these results concern both the sodium and potassium ion currents INa and IK. Specifically, Na+ channel activation has long been known to be coupled to inactivation during a depolarizing voltage-clamp step. This feature appears to be required to simulate the pulse train results in a revised model of nerve excitability. Moreover, the activation curve for IK has a significantly steeper voltage dependence, especially near its threshold (approximately -60 mV), than in the standard model, which contributes to reduced excitability, and the fully activated current-voltage relation for IK has a nonlinear, rather than a linear, dependence on driving force. An additional aspect of the revised model is accumulation/depeletion of K+ in the space between the axon and the glial cells surrounding the axon, which is significant even during a single action potential and which can account for the 15-20 mV difference between the potassium equilibrium potential EK and the maximum afterhyperpolarization of the action potential. The modifications in IK can also account for the shape of voltage changes near the foot of the action potential.

Action Potentials↗
PubMed1997

Ion conductance of the Ca(2+)-activated maxi-K+ channel from the embryonic rat brain.

By using single-channel recording techniques, we measured the conductance (gK) of the Ca(2+)-activated Maxi-K+ channel from the embryonic rat brain, and examined its dependence on K+ ions present in equimolar concentrations on both sides of the membrane patch. With ionic strength maintained constant by substitution of N-methyl-D-glucamine for K+, gK has a sigmoidal dependence upon [K+]. This result has been obscured in previous work by variations in ionic strength, which has a marked effect on single-channel conductance, especially in the limit for which this variable approaches zero. The gK versus [K+] relationship is described, theoretically, by a three-barrier, two-binding-site model in which the barrier that an ion must cross to leave the channel is decreased as [K+] is increased.

Animals↗
PubMed1996

Effects of permeant cations on K+ channel gating in nerve axons revisited.

An increase in extracellular potassium ion concentration, Ko, significantly slows the potassium channel deactivation rate in squid giant axons, as previously shown. Surprisingly, the effect does not occur in all preparations which, coupled with the voltage independence of this result in preparations in which it does occur, suggests that it is mediated at a site outside of the electric field of the channel, and that this site is accessible to potassium ions in some preparations, but not in others. In other words, the effect does not appear to be related to occupancy of the channel by potassium ions. This conclusion is supported by a four-barrier, three-binding site model of single file diffusion through the channel in which one site, at most, is unoccupied by a potassium ion (single-vacancy model). The model is consistent with current-voltage relations with various levels of Ko, and, by definition, with multiple occupancy by K+. The model predicts that occupancy of any given site is essentially independent of Ko (or Ki). The effects of extracellular Rb+ and Cs+ on gating are strongly voltage dependent, and they were observed in all preparations investigated. Consequently, the mechanism underlying these results would appear to be different from that which underlies the effect of K+ on gating. In particular, the effect of Rb+ on gating is reduced by strong hyperpolarization, which in the context of the occupancy hypothesis, is consistent with the voltage dependence of the current-voltage relation in the presence of Rb+. The primary, novel, finding in this study is that the effects of Cs+ are counterintuitive in this regard. Specifically, the slowing of channel deactivation rate by Cs+ is also reduced by hyperpolarization, similar to the Rb+ results, whereas blockade is enhanced, which is seemingly inconsistent with the concept that occupancy of the channel by Cs+ underlies the effect of this ion on gating. This result is further elucidated by barrier modeling of the current-voltage relation in the presence of Cs+.

Animals↗
PubMed1996

Effects of intracellular K+ and Rb+ on gating of embryonic rat telencephalon Ca(2+)-activated K+ channels.

We have investigated the effects of intracellular K+ and Rb+ on single-channel currents recorded from the large-conductance Ca(2+)-activated K+ (BK) channel of the embryonic rat telencephalon using the inside-out patch-clamp technique. Our novel observation concerns the effects of these ions on rapid flickering of channel openings. Specifically, flicker gating was voltage dependent, i.e., it was reduced by depolarization in the -60 to -10 mV range with equimolar concentrations of K+ ions (150 Ko+/150 Ki+). Removal of Ki+ resulted in significant flickering at all potentials in this voltage range. In other words, the voltage dependence of flicker gating was effectively eliminated by the removal of Ki+. This suggests that a K+ ion entering the channel from the intracellular medium binds, in a voltage-dependent manner, at a site that locks the flicker gate in its open position. No effects of changes in Ki+ were observed on the primary, voltage-dependent gate of the channel. The change in flickering did not cause a change in the mean burst duration, which indicates that the primary gate is stochastically independent of the flicker gate. Intracellular Rb+ can substitute for--and is even more effective than--Ki+ with regard to suppression of flickering. Substitution of Rbi+ for Ki+ also increased the mean burst duration for V > or = -30 mV. Both effects of Rbi+ were removed by membrane hyperpolarization.

Animals↗
PubMed1996

Rhinosporidiosis: three domestic cases.

Three cases of rhinosporidiosis in Americans who had not traveled abroad are reported. We believe this is the largest cluster of indigenous cases reported in the United States. The three patients had lived in rural northeast Georgia all of their lives. One had a polypoid conjunctival lesion, and the two others had nasal polyps. In each case, the diagnosis was made by demonstrating morphologically distinctive fungal elements in histopathologic sections. Clinically, rhinosporidiosis had not been suspected.

Adolescent↗
PubMed1996

Nifedipine-associated gingival overgrowth: a survey of the literature and report of four cases.

A survey of the literature indicates that calcium channel-blocking medications are used for an ever-increasing number of medical problems. Their use may result in gingival overgrowth that can be of concern to the patient and dentist. Four cases are presented that illustrate several relevant points: (1) histologic examination may reveal factors overlooked in the medical history; (2) the condition exists in a variety of clinical manifestations; (3) the level of plaque control maintained by the patient is important to the management of the condition; and (4) responses vary to different approaches to treatment, including changes in medication, as well as nonsurgical and surgical therapies.

Adult↗
PubMed1995

A simple model of K+ channel activation in nerve membrane.

A model is proposed for activation of potassium ion channel current, IK, in squid giant axons, which consists of two closed states and one open state. The rate parameter in the forward direction between the two closed states depends upon previous history. That is, it relaxes exponentially to its steady-state value appropriate to the membrane potential of a voltage clamp step rather than change instantaneously as in traditional models of channel gating. The model successfully describes both the enhancement of the delay in activation of IK with relatively negative prepulse potentials, i.e. the Cole-Moore effect, and the time-dependent rising phase of "on" gating current, which has been reported recently for several types of potassium channels.

Animals↗
PubMed1995

Quaternary ammonium ion blockade of IK in nerve axons revisited. Open channel block vs. state independent block.

The mechanism of blockade of the delayed rectifier potassium ion channel in squid giant axons by intracellular quaternary ammonium ions (QA) appears to be remarkably sensitive to the structure of the blocker. TEA, propyltriethyl-ammonium (C3), and propyltetraethylammonium (TAA-C3) all fail to alter the deactivation, or "tail" current time course following membrane depolarization, even with relatively large concentrations of the blockers, whereas butyltriethylammonium (C4), butyltetraethylammonium (TAA-C4), and pentytriethyammonium (C5) clearly do have such an effect. The relative electrical distance of blockade for all of these ions is approximately 0.25-0.3 from the inner surface of the membrane. The observations concerning TEA, C3, and TAA-C3 suggest that these ions can block the channel in either its open or its closed state. The results with C4, TAA-C4, and C5 are consistent with the open channel block model. Moreover, the sensitivity of block mechanism to the structure of the blocker suggests that the gate is located close to the QA ion binding site and that TEA, C3, and TAA-C3 do not interfere with channel gating, whereas C4, TAA-C4, C5, and ions having a longer hydrophobic "tail" than C5 do have such an effect. The parameters of block obtained for all QA ions investigated were unaffected by changes in the extracellular potassium ion concentration.

Animals↗