Odorants suppress a voltage-activated K+ conductance in rat olfactory neurons.
(49/1319)
Stimulation of olfactory receptor neurons (ORNs) with odors elicits an increase in the concentration of cAMP leading to opening of cyclic nucleotide-gated (CNG) channels and subsequent depolarization. Although opening of CNG channels is thought to be the main mechanism mediating signal transduction, modulation of other ion conductances by odorants has been postulated. To determine whether K+ conductances are modulated by odorants in mammalian ORNs, we examined the response of rat ORNs to odors by recording membrane current under perforated-patch conditions. We find that rat ORNs display two predominant types of responses. Thirty percent of the cells responded to odorants with activation of a CNG conductance. In contrast, in 55% of the ORNs, stimulation with odorants inhibited a voltage-activated K+ conductance (IKo). In terms of pharmacology, ion permeation, outward rectification, and time course for inactivation, IKo resembled a delayed rectifier K+ conductance. The effect of odorants on IKo was specific (only certain odorants inhibited IKo in each ORN) and concentration dependent, and there was a significant latency between arrival of odorants to the cell and the onset of suppression. These results indicate that indirect suppression of a K+ conductance (IKo) by odorants plays a role in signal transduction in mammalian ORNs. (+info)
Dendrodendritic recurrent excitation in mitral cells of the rat olfactory bulb.
(50/1319)
Most neuronal interactions within the olfactory bulb network are mediated by dendrodendritic synapses. Dendritic transmitter release potentially could affect the parent dendrite as well as local neuronal elements that have receptors for the released transmitter. Here we report that under conditions that facilitate N-methyl-D-aspartate (NMDA) receptor activity (reduced GABAA inhibition and extracellular Mg2+), a single action potential evoked by brief intracellular current pulses in mitral cells is followed by a prolonged depolarization, which is blocked by an NMDA receptor antagonist. This depolarization also is evoked by a presumed calcium spike in the presence of tetrodotoxin. A similar NMDA-receptor-dependent prolonged depolarization is elicited by stimulation of the lateral olfactory tract at current intensities subthreshold for antidromic activation of the recorded neuron. These observations suggest that glutamate released from the dendrites of mitral cells excites the same and neighboring mitral cell dendrites. Further evidence suggests that both the apical and lateral dendrites of mitral cells participate in this recurrent excitation. These dendrodendritic interactions may play a role in the prolonged, NMDA-receptor-dependent depolarization of mitral/tufted cells evoked by olfactory nerve stimulation. (+info)
Glutamate receptors mediate TTX-resistant synchronous activity in the rat hippocampus.
(51/1319)
4-Aminopyridine (4-AP) is a well known convulsant that enhances the release of both excitatory and inhibitory neurotransmitters in the CNS. Low concentrations of 4-AP (approximately 100 microM) readily induce synchronized discharges in the hippocampus that are blocked by tetrodotoxin (TTX), suggesting that they require Na(+)-dependent action potentials in addition to the enhanced release of neurotransmitters. However, in the present study we have found that higher concentrations of 4-AP (1 mM) in combination with 5 mM tetraethylammonium (TEA) induce spontaneous synchronized discharges in rat hippocampal slices that are resistant to blockade by TTX. These synchronous discharges are evident in field potential recordings, which progress from the hilus to CA1 at 0.023 +/- 0.002 m/sec and in intracellular recordings from the hilar mossy cells and CA3 pyramidal cells. In some slices exposed to 4-AP and TEA, smaller-amplitude asynchronous responses also were recorded. 4-AP-induced spontaneous discharges are blocked by 20 microM DNQX and by 100 microM Cd(2+) but are resistant to blockade by either 25 microM bicuculline or 25 microM D-APV. These results suggest that the activation of postsynaptic AMPA receptors is necessary to produce TTX-resistant synchronized discharges. The laminar profile of field potentials recorded in CA3 and CA1 suggests that glutamate is released from axons of CA3 pyramidal cells despite the blockade of fast axonal Na(+) channels by TTX. Synchronous discharges may result from glutamate released at proximal recurrent collaterals after spontaneous Ca(2+) spikes in CA3 pyramidal cells. (+info)
Determinants of excitability at transition zones in Kv1.1-deficient myelinated nerves.
(52/1319)
This study examines the role of K channel segregation and fiber geometry at transition zones of mammalian nerve terminals in the peripheral nervous system. Mutant mice that are deficient in Kv1.1, a fast Shaker K channel normally localized beneath the myelin sheath, display three types of cooling-induced abnormal hyperexcitability localized to regions before the transition zones of myelinated nerves. The first type is stimulus-evoked nerve backfiring that is absent at birth, peaks at postnatal day 17 (P17), and subsides in adults. The second type is spontaneous activity that has a more delayed onset, peaks at P30, and also disappears in older mice (>P60). TEA greatly amplifies this spontaneous activity with an effective dosage of approximately 0.7 mM, and can induce its reappearance in older mutant mice (>P100). These first two types of hyperexcitability occur only in homozygous mutants that are completely devoid of Kv1.1. The third type occurs in heterozygotes and represents a synergism between a TEA-sensitive channel and Kv1.1. Heterozygotes exposed to TEA display no overt phenotype until a single stimulation is given, which is then followed by an indefinite phase of repetitive discharge. Computer modeling suggests that the excitability of the transition zone near the nerve terminal has at least two major determinants: the preterminal internodal shortening and axonal slow K channels. We suggest that variations in fiber geometry create sites of inherent instability that is normally stabilized by a synergism between myelin-concealed Kv1.1 and a slow, TEA-sensitive K channel. (+info)
Modulation of glioma cell migration and invasion using Cl(-) and K(+) ion channel blockers.
(53/1319)
Human malignant gliomas are highly invasive tumors. Mechanisms that allow glioma cells to disseminate, migrating through the narrow extracellular brain spaces are poorly understood. We recently demonstrated expression of large voltage-dependent chloride (Cl(-)) currents, selectively expressed by human glioma cells in vitro and in situ (Ullrich et al., 1998). Currents are sensitive to several Cl(-) channel blockers, including chlorotoxin (Ctx), (Ullrich and Sontheimer; 1996; Ullrich et al; 1996), tetraethylammonium chloride (TEA), and tamoxifen (Ransom and Sontheimer, 1998). Using Transwell migration assays, we show that blockade of glioma Cl(-) channels specifically inhibits tumor cell migration in a dose-dependent manner. Ctx (5 microM), tamoxifen (10 microM), and TEA (1 mM) also prevented invasion of human glioma cells into fetal rat brain aggregates, used as an in vitro model to assess tumor invasiveness. Anion replacement studies suggest that permeation of chloride ions through glioma chloride channel is obligatory for cell migration. Osmotically induced cell swelling and subsequent regulatory volume decrease (RVD) in cultured glioma cells were reversibly prevented by 1 mM TEA, 10 microM tamoxifen, and irreversibly blocked by 5 microM Ctx added to the hypotonic media. Cl(-) fluxes associated with adaptive shape changes elicited by cell swelling and RVD in glioma cells were inhibited by 5 microM Ctx, 10 microM tamoxifen, and 1 mM TEA, as determined using the Cl(-)-sensitive fluorescent dye 6-methoxy-N-ethylquinolinium iodide. Collectively, these data suggest that chloride channels in glioma cells may enable tumor invasiveness, presumably by facilitating cell shape and cell volume changes that are more conducive to migration and invasion. (+info)
Voltage-dependent outward K(+) current in intermediate cell of stria vascularis of gerbil cochlea.
(54/1319)
A voltage-dependent outward K(+) (K(V)) current in the intermediate cell (melanocyte) of the cochlear stria vascularis was studied using the whole cell patch-clamp technique. The K(V) current had an activation threshold voltage of approximately -80 mV, and 50% activation was observed at -42.6 mV. The time courses of activation and inactivation were well fitted by two exponential functions: the time constants at 0 mV were 7.9 and 58.8 ms for activation and 0.6 and 4.3 s for inactivation. The half-maximal activation time was 13. 8 ms at 0 mV. Inactivation of the current was incomplete even after a prolonged depolarization of 10 s. This current was independent of intracellular Ca(2+). Quinine, verapamil, Ba(2+), and tetraethylammonium inhibited the current in a dose-dependent manner, but 4-aminopyridine was ineffective at 50 mM. We conclude that the K(V) conductance in the intermediate cell may stabilize the membrane potential, which is thought to be closely related to the endocochlear potential, and may provide an additional route for K(+) secretion into the intercellular space. (+info)
Four different components contribute to outward current in rat ventricular myocytes.
(55/1319)
In rat ventricle, two Ca(2+)-insensitive components of K(+) current have been distinguished kinetically and pharmacologically, the transient, 4-aminopyridine (4-AP)-sensitive I(to) and the sustained, tetraethylammonium (TEA)-sensitive I(K). However, a much greater diversity of depolarization-activated K(+) channels has been reported on the level of mRNA and protein. In the search for electrophysiological evidence of further current components, the whole cell voltage-clamp technique was used to analyze steady-state inactivation of outward currents by conditioning potentials in a wide voltage range. Peak (I(peak)) and late (I(late)) currents during the test pulse were analyzed by Boltzmann curve fitting, producing three fractions each. Fractions a and b had different potentials of half-maximum inactivation (V(0.5)); the third residual fraction, r, did not inactivate. Fractions a for I(peak) and I(late) had similar relative amplitudes and V(0.5) values, whereas size and V(0.5) of fractions b differed significantly between I(peak) and I(late). Only b of I(peak) was transient, suggesting a relation with I(to), whereas a, b, and r of I(late) appeared to be three different sustained currents. Therefore, four individual outward current components were distinguished: I(to) (b of I(peak)), I(K) (a), the steady-state current I(ss) (r), and the novel current I(Kx) (b of I(late)). This was further supported by differential sensitivity to TEA, 4-AP, clofilium, quinidine, dendrotoxin, heteropodatoxin, and hanatoxin. With the exception of I(to), none of the currents exhibited a marked transmural gradient. Availability of I(K) was low at resting potential; nevertheless, I(K) contributed to action potential shortening in hyperpolarized subendocardial myocytes. In conclusion, on the basis of electrophysiological and pharmacological evidence, at least four components contribute to outward current in rat ventricular myocytes. (+info)
Delayed rectifier currents in rat globus pallidus neurons are attributable to Kv2.1 and Kv3.1/3.2 K(+) channels.
(56/1319)
The symptoms of Parkinson disease are thought to result in part from increased burst activity in globus pallidus neurons. To gain a better understanding of the factors governing this activity, we studied delayed rectifier K(+) conductances in acutely isolated rat globus pallidus (GP) neurons, using whole-cell voltage-clamp and single-cell RT-PCR techniques. From a holding potential of -40 mV, depolarizing voltage steps in identified GP neurons evoked slowly inactivating K(+) currents. Analysis of the tail currents revealed rapidly and slowly deactivating currents of similar amplitude. The fast component of the current deactivated with a time constant of 11. 1 +/- 0.8 msec at -40 mV and was blocked by micromolar concentrations of 4-AP and TEA (K(D) approximately 140 microM). The slow component of the current deactivated with a time constant of 89 +/- 10 microseconds at -40 mV and was less sensitive to TEA (K(D) = 0.8 mM) and 4-AP (K(D) approximately 6 mM). Organic antagonists of Kv1 family channels had little or no effect on somatic currents. These properties are consistent with the hypothesis that the rapidly deactivating current is attributable to Kv3.1/3.2 channels and the slowly deactivating current to Kv2.1-containing channels. Semiquantitative single-cell RT-PCR analysis of Kv3 and Kv2 family mRNAs supported this conclusion. An alteration in the balance of these two channel types could underlie the emergence of burst firing after dopamine-depleting lesions. (+info)