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S R Levinson

Publications and source records attributed to S R Levinson.

At least 19 recordsLinked to original sources
PubMed1995

Voltage- and frequency-dependent pentobarbital suppression of brain and muscle sodium channels expressed in a mammalian cell line.

The voltage- and frequency-dependent interactions of pentobarbital with voltage-gated sodium channels were examined in whole-cell patch-clamp recordings. Using rat brain IIA and rat muscle rSkM1 sodium channels expressed in stably transfected Chinese hamster ovary cell lines, it was found that pentobarbital reduced peak inward sodium currents with IC50 values of 1.2 mM (brain) and 1.0 mM (muscle). Analysis of steady state channel availability curves revealed two distinct effects of pentobarbital on both channel isoforms, i.e., a voltage-independent current reduction and an additional hyperpolarizing shift in the voltage dependence of channel availability. The latter effect leads to a voltage dependence of pentobarbital potency. Pentobarbital was also found to slow channel recovery after depolarization, yielding an additional use-dependent component of current suppression. Use-dependent block was enhanced by higher stimulation frequencies, longer pulse durations, and more depolarized holding and pulse potentials. All effects were identical for both channels. These findings can be explained in terms of the modulated receptor hypothesis and are consistent with a preferential interaction of pentobarbital with the inactivated channel state. As a consequence, actual pentobarbital potency would depend largely on experimental conditions or, in vivo, on the physiological parameters of a particular cell.

Animals↗
PubMed1995

Clustering of Na+ channels and node of Ranvier formation in remyelinating axons.

Polyclonal antibodies were raised against a well conserved region of the vertebrate Na+ channel and were affinity purified for use in immunocytochemistry. Focal demyelination of rat sciatic axons was initiated by an intraneural injection of lysolecithin and Na+ channel clustering was followed at several stages of myelin removal and repair. At 1 week post-injection axons contained long, fully demyelinated regions. Na+ channel clusters appeared only at heminodes forming the borders of these zones, and at widely spaced isolated sites that may represent former nodes of Ranvier. Over the next few days proliferating Schwann cells adhered to axons and began to extend processes. Clusters of Na+ channels appeared at the edges of these structures. As the Schwann cells elongated, the clusters seemed to move with them, since they remained at edges and the distance between aggregates increased. Clusters associated with different Schwann cells ultimately approached each other and appeared to fuse. Na+ channels then coalesced further at these sites, forming new nodes of Ranvier in regions that previously were internodal. If Schwann cell proliferation were blocked by mitomycin, no new clusters of Na+ channels appeared within internodes. Under these conditions, heminodal clusters remained visible at 1 week postinjection, but by 2 weeks they were no longer detectable, suggesting that proliferating Schwann cells are required for their maintenance. Clusters at normal nodes of Ranvier remained. It is concluded that Na+ channel aggregation and mobility in demyelinated nerve fibers is controlled by adhering Schwann cells, resulting in the formation of stable new nodes of Ranvier during remyelination.

Animals↗
PubMed1994

Sodium channels accumulate at the tips of injured axons.

The axolemmal distribution of voltage-gated sodium channels largely determines the regions of axonal electrical excitability. Using a well-characterized anti-sodium channel antibody, we examined peripheral nerve fibers focally injured by exposure to the neurotoxic agent, potassium tellurite (K2TeO3). Immunocytochemical and radioimmunoassay data showed a focal accumulation of sodium channels within the tips of injured axons. The major increase in sodium channel concentration occurred between 7 and 11 days after toxin exposure; however, immunocytochemically, excess sodium channels persisted in several axonal endings for a much longer time. The accumulation of sodium channels at injured axonal tips may be responsible, in part, for ectopic axonal excitability and the resulting abnormal sensory phenomena (especially pain and paresthesias) which frequently complicate peripheral nerve injury in humans.

Animals↗
PubMed1994

TTX-sensitive dendritic sodium channels underlie oscillatory discharge in a vertebrate sensory neuron.

Immunocytochemical and electrophysiological techniques were used to localize TTX-sensitive sodium channels (NaChs) over the soma-dendritic axis of basilar and nonbasilar pyramidal cells of the electrosensory lateral line lobe (ELL) of weakly electric fish (Apteronotus leptorhynchus). Dense NaCh-like immunolabel was detected on the membranes of basilar and nonbasilar pyramidal cell somata. Punctate regions of immunolabel (approximately 15 microns) were separated by nonlabeled expanses of membrane over the entire extent of basal dendrites. Similar punctate immunolabel was observed over the apical dendrites, and frequently on membranes of afferent parallel fiber boutons in the distal apical dendritic region. Intracellular recordings from pyramidal cell somata or proximal apical dendrites (75-200 microns) were obtained using an in vitro ELL slice preparation. TTX-sensitive potentials were identified by focal pressure ejection of TTX. Somatic recordings demonstrated both TTX-sensitive fast spike discharge and a slow prepotential; similar but lower amplitude potentials were recorded in apical dendrites. Dendritic spikes were composed of at least two active components triggered by a fast prepotential (FPP) generated by the somatic spike. TTX-sensitive spikes propagated in a retrograde fashion over at least the proximal 200 microns of the apical dendrites, as determined by the conduction of an antidromic population spike and focal TTX ejections. Somatic spikes were followed by a depolarizing afterpotential (DAP) that was similar in duration and refractory period to that of proximal dendritic spikes. During repetitive spike discharge, the DAP could increase in amplitude and attain somatic spike threshold, generating a high-frequency spike doublet and a subsequent hyperpolarization that terminated spike discharge. Repetition of this process gave rise to an oscillatory burst discharge (2-6 spikes/burst) with a frequency of 40-80 Hz. Both the DAP and oscillatory discharge were selectively blocked by TTX ejections restricted to the proximal apical dendritic region. The present study demonstrates an immunolocalization of NaChs over somatic and dendritic membranes of a vertebrate sensory neuron that correlates with the distribution of TTX-sensitive potentials. The interaction of somatic and dendritic action potentials is further shown to underlie an oscillatory discharge believed to be important in electrosensory processing.

Animals↗
PubMed1992

Grayanotoxin-I-modified eel electroplax sodium channels. Correlation with batrachotoxin and veratridine modifications.

To probe the structure-function relationships of voltage-dependent sodium channels, we have been examining the mechanisms of channel modification by batrachotoxin (BTX), veratridine (VTD), and grayanotoxin-I (GTX), investigating the unifying mechanisms that underlie the diverse modifications of this class of neurotoxins. In this paper, highly purified sodium channel polypeptides from the electric organ of the electric eel were incorporated into planar lipid bilayers in the presence of GTX for comparison with our previous studies of BTX (Recio-Pinto, E., D. S. Duch, S. R. Levinson, and B. W. Urban. 1987. J. Gen. Physiol. 90:375-395) and VTD (Duch, D. S., E. Recio-Pinto, C. Frenkel, S. R. Levinson, and B. W. Urban. 1989. J. Gen. Physiol. 94:813-831) modifications. GTX-modified channels had a single channel conductance of 16 pS. An additional large GTX-modified open state (40-55 pS) was found which occurred in bursts correlated with channel openings and closings. Two voltage-dependent processes controlling the open time of these modified channels were characterized: (a) a concentration-dependent removal of inactivation analogous to VTD-modified channels, and (b) activation gating similar to BTX-modified channels, but occurring at more hyperpolarized potentials. The voltage dependence of removal of inactivation correlated with parallel voltage-dependent changes in the estimated K1/2 of VTD and GTX modifications. Ranking either the single channel conductances or the depolarization required for 50% activation, the same sequence is obtained: unmodified > BTX > GTX > VTD. The efficacy of the toxins as activators follows the same ranking (Catterall, W. A. 1977. J. Biol. Chem. 252:8669-8676).

Animals↗
PubMed1991

Increased numbers of sodium channels form along demyelinated axons.

Sodium channels, which are largely localized to the nodes of Ranvier in myelinated axons, appear to form new distributions along demyelinated axons. In this study a sensitive radioimmunoassay (RIA) was used to examine the changes in the total number of sodium channels that occur in nerves experimentally demyelinated in vivo with doxorubicin (adriamycin). The results clearly illustrate the development of an increased number of sodium channels during demyelination, suggesting that this process is associated with the formation of new sodium channels.

Animals↗
PubMed1991

Monoclonal antibodies raised against post-translational domains of the electroplax sodium channel.

Eleven monoclonal antibodies were identified that recognized eel electroplax sodium channels. All the monoclonal antibodies specifically immunostained the mature TTX-sensitive sodium channel (Mr 265,000) on immunoblots. None of the monoclonal antibodies would precipitate the in vitro translated channel core polypeptide in solution. One monoclonal antibody, 3G4, was found to bind to an epitope involving terminal polysialic acids. Extensive digestion of the channel by the exosialidase, neuraminidase, or partial polysialic acid removal by the endosialidase, endo-N-acetylneuraminidase, destroy the 3G4 epitope. 3G4 is, therefore, a highly selective probe for the post-translationally attached polysialic acids. Except for this monoclonal antibody, the epitopes recognized by the remaining antibodies were highly resistant to extensive N-linked deglycosylation. Thus, the monoclonal antibodies may be directed against unique post-translationally produced domains of the electroplax sodium channel, presumably sugar groups that are abundant on this protein (Miller, J.A., Agnew, W.S., Levinson, S.R. 1983, Biochemistry 22:462-470). These monoclonal antibodies should prove useful as tools to study discrete post-translational processing events in sodium channel biosynthesis.

Animals↗
PubMed1991

Diagnostic nasal endoscopy and functional endoscopic sinus surgery: an update and review of complications.

Diagnostic nasal endoscopy and functional endoscopic sinus surgery are relatively new techniques that have expanded our understanding of sinus physiology and the etiologies of sinus pathology. Diagnosis at an early stage of chronic sinus disease demonstrates that pathological changes are often limited to the osteomeatal complex and the anterior sinus group. Early disease refractory to aggressive medical management usually responds to surgical treatment. Surgery in this setting is most often performed under local anesthesia, without nasal packing, with an immediate improvement in the patient's symptoms, and with minimal risk of complications.

Chronic Disease↗
PubMed1990

Neuraminidase treatment modifies the function of electroplax sodium channels in planar lipid bilayers.

Sodium channels from several sources are covalently modified by unusually large numbers of negatively charged sialic acid residues. In the present studies, purified electroplax sodium channels were treated with neuraminidase to remove sialic acid residues and then examined for functional changes in planar lipid bilayers. Neuraminidase treatment resulted in a large depolarizing shift in the average potential required for channel activation. Additionally, desialidated channels showed a striking increase in the frequency of reversible transitions to subconductance states. Thus it appears that sialic acid residues play a significant role in the function of sodium channels, possibly through their influence on the local electric field and/or conformational stability of the channel molecule.

Animals↗
PubMed1990

Changed distribution of sodium channels along demyelinated axons.

Voltage-gated sodium channels are largely localized to the nodes of Ranvier in myelinated axons, providing a physiological basis for saltatory conduction. What happens to these channels in demyelinated axons is not known with certainty. Experimentally demyelinated axons were examined by using a well-characterized, polyclonal antibody directed against sodium channels. Immunocytochemical and radioimmunoassay data were consistent with the distribution of an increased number of sodium channels along segments of previously internodal axon. These findings affirm the plasticity of sodium channels in demyelinated axolemma and may be relevant to understanding how axons recover conduction after demyelination.

Animals↗
PubMed1989

Na+ channel accumulation on axolemma of afferent endings in nerve end neuromas in Apteronotus.

In mammals, cut sensory axons trapped in a nerve end neuroma have been shown to develop hyperexcitability, and to become a source of ectopic afferent discharge and abnormal sensation. We have explored cellular mechanisms underlying neuroma electrogenesis. First we confirmed that ectopic neuroma discharge develops in injured afferents in the electrosensory lateral line nerve of the weakly electric fish Apteronotus, as it does in mammals. Then, using previously characterized antibodies that specifically recognize Na+ channel proteins in this species, we obtained light and electron microscopic evidence of abnormally intense immunolabelling of axolemma at the injury site. Accumulation of excess Na+ channels in afferent endings in neuromas could account for their electrical hyperexcitability.

Animals↗
PubMed1989

Veratridine modification of the purified sodium channel alpha-polypeptide from eel electroplax.

In the interest of continuing structure-function studies, highly purified sodium channel preparations from the eel electroplax were incorporated into planar lipid bilayers in the presence of veratridine. This lipoglycoprotein originates from muscle-derived tissue and consists of a single polypeptide. In this study it is shown to have properties analogous to sodium channels from another muscle tissue (Garber, S. S., and C. Miller. 1987. Journal of General Physiology. 89:459-480), which have an additional protein subunit. However, significant qualitative and quantitative differences were noted. Comparison of veratridine-modified with batrachotoxin-modified eel sodium channels revealed common properties. Tetrodotoxin blocked the channels in a voltage-dependent manner indistinguishable from that found for batrachotoxin-modified channels. Veratridine-modified channels exhibited a range of single-channel conductance and subconductance states. The selectivity of the veratridine-modified sodium channels for sodium vs. potassium ranged from 6-8 in reversal potential measurements, while conductance ratios ranged from 12-15. This is similar to BTX-modified eel channels, though the latter show a predominant single-channel conductance twice as large. In contrast to batrachotoxin-modified channels, the fractional open times of these channels had a shallow voltage dependence which, however, was similar to that of the slow interaction between veratridine and sodium channels in voltage-clamped biological membranes. Implications for sodium channel structure are discussed.

Animals↗
PubMed1987

Biosynthesis of electroplax sodium channels in Electrophorus electrocytes and Xenopus oocytes.

We have synthesized the eel electroplax sodium channel core polypeptide in both a cell-free and a frog oocyte system and report it does not possess the unusual electrophoretic properties of the mature, native sodium channel polypeptide isolated from electroplax membranes. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the mature channel polypeptide exhibits both a diffuse banding pattern (microheterogeneity) and an extremely high electrophoretic free mobility. In contrast, the core polypeptide synthesized in vitro or in vivo migrates as a sharp band with a near-normal electrophoretic free mobility (Mr 230,000). The microheterogeneity of the mature peptide has been inferred to result from varying degrees of glycosylation of the channel polypeptide [Miller, J.A., Agnew, W.S., & Levinson, S.R. (1983) Biochemistry 22, 462-470]. We present evidence here that the anomalously high electrophoretic free mobility is due to the binding of large amounts of sodium dodecyl sulfate to posttranslationally modified domains on the protein. In addition, we have followed the posttranslational processing of eel sodium channels in both the eel electrocyte and the frog oocyte. Using lectin binding and Ferguson analysis, we found that the channel was processed relatively rapidly to an intermediate form in the Golgi apparatus that apparently contained fewer carbohydrate and hydrophobic domains than the mature channel. The further addition of carbohydrate and hydrophobic domains, which are required before the channel acquires its characteristic physicochemical properties, proceeded relatively slowly in the electrocyte and appeared not to have occurred to the majority of intermediately processed channels in the frog oocyte.

Animals↗
PubMed1987

Distribution of (Na+ + K+)ATPase and sodium channels in skeletal muscle and electroplax.

The distributions of (Na+ + K+)ATPase and sodium channels in skeletal muscle fibres and electrocytes were determined by immunofluorescent and immunoelectron microscopic techniques using antibodies against rat and eel (Na+ + K+)ATPase and the eel electric organ sodium channel. The extrajunctional sarcolemma of skeletal muscle was uniformly stained by polyclonal antibodies against (Na+ + K+)ATPase and the sodium channel. The T-tubule system of skeletal muscle was also labelled heavily for both (Na+ + K+)ATPase and the sodium channel. The terminal cisternae of the sarcoplasmic reticulum was stained for (Na+ + K+)ATPase but not sodium channels. At the motor endplate, (Na+ + K+)ATPase-like immunoreactivity was present along the plasmalemma of motor nerve terminals but not along the postsynaptic junctional sarcolemma. Paradoxically, a monoclonal antibody that binds to the alpha form of the catalytic subunit of (Na+ + K+)ATPase from rat hepatocytes and renal tubule cells did not label the enzyme in rat skeletal muscle. In electrocytes, (Na+ + K+)ATPase-like immunoreactivity was concentrated primarily along the plasmalemma and calveolae of the non-innervated face. In contrast, sodium channel-like immunoreactivity was concentrated along the plasmalemma of the innervated face except in the clefts of the postsynaptic membrane. Thus, we conclude that at endplates both the (Na+ + K+)ATPase of rat skeletal muscle and sodium channels of eel electrocytes are not concentrated in the juxtaneuronal postsynaptic membrane. We also interpret the failure of the monoclonal anti-alpha (Na+ + K+)ATPase antibodies to bind to the enzyme in muscle to indicate that the catalytic subunit of skeletal muscle (Na+ + K+)ATPase displays different epitopes than does the alpha subunit of kidney and liver.

Animals↗
PubMed1987

Neurotoxin-modulated uptake of sodium by highly purified preparations of the electroplax tetrodotoxin-binding glycopeptide reconstituted into lipid vesicles.

Using the dialysable detergent CHAPS (3-[(3-cholamidopropyl)-dimethylammonio]-1-propane sulfonate), the tetrodotoxin-binding protein from the electroplax of the electric eel has been purified to a high degree of both chemical homogeneity and toxin-binding activity. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the best preparations showed only a single microheterogeneous band at Mr approximately 260,000, despite attempts to visualize smaller bands by sample overloading. Upon dialysis, this material became incorporated into the membranes of small unilamellar vesicles, and in this form the purified protein exhibited tetrodotoxin-binding properties similar to the component in the original electroplax membrane. Furthermore, in the presence of activator neurotoxins the vesicles were able to accumulate isotopic sodium in a manner similar to that previously described for less active or less pure preparations of vesicles containing either mammalian or eel electroplax toxin-binding proteins. Quantitative consideration of the isotopic transport activity of this pure material, along with the high degree of purity of the protein, strongly suggests that the 260-kDa glycopeptide from electroplax is necessary and sufficient to account for the sodium channel function seen in these studies, and eliminates the possible involvement of smaller peptides in the channel phenomena observed.

Animals↗