Tetanus toxin: convulsant action on mouse spinal cord neurons in culture

Bergey, G. K.; Macdonald, R. L.; Habig, William H.; Hardegree, M. C.; Nelson, P. G.

doi:10.1523/jneurosci.03-11-02310.1983

Cited by 66 publications

(56 citation statements)

References 52 publications

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“…In particular, it is possible that there were slow PSPs that we did not detect, such as those produced by metabotropic glutamate receptors or neuromodulators. For this reason, an alternative and independent method was used to block synaptic transmission, tetanus toxin, which blocks neurotransmitter release (Bergey et al 1983). Tetanus toxin also circumvents the potential problem that some receptor antagonists have been reported to act as weak agonists for excitatory amino acid and neuromodulator receptors (Collingridge and Lester 1989;Colquhoun and Patrick 1997).…”

Section: Cultures Were Active In the Presence Of Tetanus Toxinmentioning

confidence: 99%

Intrinsic Dynamics in Neuronal Networks. II. Experiment

Latham

Richmond

Nirenberg

et al. 2000

Journal of Neurophysiology

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Intrinsic dynamics in neuronal networks. II. Experiment. J. Neurophysiol. 83: 828 -835, 2000. Neurons in many regions of the mammalian CNS remain active in the absence of stimuli. This activity falls into two main patterns: steady firing at low rates and rhythmic bursting. How these firing patterns are maintained in the presence of powerful recurrent excitation, and how networks switch between them, is not well understood. In the previous paper, we addressed these issues theoretically; in this paper we address them experimentally. We found in both studies that a key parameter in controlling firing patterns is the fraction of endogenously active cells. The theoretical analysis indicated that steady firing rates are possible only when the fraction of endogenously active cells is above some threshold, that there is a transition to bursting when it falls below that threshold, and that networks becomes silent when the fraction drops to zero. Experimentally, we found that all steadily firing cultures contain endogenously active cells, and that reducing the fraction of such cells in steadily firing cultures causes a transition to bursting. The latter finding implies indirectly that the elimination of endogenously active cells would cause a permanent drop to zero firing rate. The experiments described here thus corroborate the theoretical analysis. I N T R O D U C T I O NMany areas of the mammalian CNS exhibit neuronal activity in the absence of stimuli. Two patterns of activity that are commonly observed are steady firing at low rates and rhythmic bursting. How do dynamic interactions between excitatory and inhibitory neurons produce these firing patterns? Specifically, how do networks remain stable in the face of powerful recurrent excitation, and how do they switch between steady firing and bursting? In the previous paper (Latham et al. 2000) we investigated these questions theoretically. We found that firing patterns are controlled largely by one parameter, the fraction of endogenously active cells; i.e., the fraction of cells that fire without input. When no endogenously active cells are present, networks are either silent or fire at a high rate; as the number of endogenously active cells increases, there is a transition to bursting; and with a further increase, there is a second transition to steady firing at a low rate. The main assumptions in our theoretical analysis were conventional: excitatory input to a neuron increases its firing rate, inhibitory input decreases it, and neurons exhibit spike-frequency adaptation.In this paper we explore experimentally the link between endogenous activity and intrinsic firing patterns. To achieve the control necessary to effectively test the theoretical predictions, we used cultured mouse spinal cord neurons. Such networks are easily accessible, are amenable to pharmacological intervention, and are consistent with critical model assumptions, including the existence of spike-frequency adaptation (Kernell 1965(Kernell , 1972Tseng and Prince 1993) and the expression of both exc...

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Section: Cultures Were Active In the Presence Of Tetanus Toxinmentioning

confidence: 99%

Intrinsic Dynamics in Neuronal Networks. II. Experiment

Latham

Richmond

Nirenberg

et al. 2000

Journal of Neurophysiology

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“…toxin action is manifest initially as a loss of inhibitory postsynaptic potentials and the onset of convulsant activity (23,24). In a previous study, we examined the effect of TeNT on potassium-evoked neurotransmitter release in murine spinal cord cell cultures (25) and demonstrated that inhibitory glycinergic neurons are particularly sensitive to the toxin.…”

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confidence: 99%

Neuronal Sensitivity to Tetanus Toxin Requires Gangliosides

Williamson¹,

Bateman²,

Clifford³

et al. 1999

Journal of Biological Chemistry

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Tetanus toxin produces spastic paralysis in situ by blocking inhibitory neurotransmitter release in the spinal cord. Although di-and trisialogangliosides bind tetanus toxin, their role as productive toxin receptors remains unclear. We examined toxin binding and action in spinal cord cell cultures grown in the presence of fumonisin B 1 , an inhibitor of ganglioside synthesis. Mouse spinal cord neurons grown for 3 weeks in culture in 20 M fumonisin B 1 develop dendrites, axons, and synaptic terminals similar to untreated neurons, even though thin layer chromatography shows a greater than 90% inhibition of ganglioside synthesis. Absence of tetanus and cholera toxin binding by toxin-horseradish peroxidase conjugates or immunofluorescence further indicates loss of mono-and polysialogangliosides. In contrast to control cultures, tetanus toxin added to fumonisin B 1 -treated cultures does not block potassiumstimulated glycine release, inhibit activity-dependent uptake of FM1-43, or abolish immunoreactivity for vesicle-associated membrane protein, the toxin substrate. Supplementing fumonisin B 1 -treated cultures with mixed brain gangliosides completely restores the ability of tetanus toxin to bind to the neuronal surface and to block neurotransmitter release. These data demonstrate that fumonisin B 1 protects against toxin-induced synaptic blockade and that gangliosides are a necessary component of the receptor mechanism for tetanus toxin.Tetanus toxin (TeNT) 1 blocks the release of inhibitory neurotransmitters in the spinal cord leading to hyperactivity of motor neurons and consequent spastic paralysis (for review, see Ref. 1). The toxin is synthesized as a single polypeptide (M r ϭ 150,000), released by bacterial lysis, and cleaved to form an active dichain toxin with the heavy and light chains linked through one disulfide bond. The carboxyl-terminal half of the heavy chain constitutes the receptor binding domain, the amino-terminal half of the heavy chain is responsible for membrane translocation of the toxin, and the light chain contains the catalytic domain (for review, see Ref.2). Upon entering the synaptic cytosol, the toxin arrests neurotransmitter release by its action as a zinc endopeptidase, which cleaves vesicle-associated membrane protein (VAMP), a synaptic vesicle integral membrane protein believed to be critical for neuroexocytosis (3, 4). Whereas TeNT has been shown to bind to the surface of neurons and to act intracellularly (for reviews, see Refs. 2 and 5-7), its cell surface receptor(s) and mechanism of delivery to the cytosol are not well understood.Since the demonstration of the affinity of TeNT for gangliosides present on the neuronal surface (8), there have been a number of studies (for review, see Ref. 6) characterizing the binding of TeNT to gangliosides in various in vitro and in vivo preparations. Although it is difficult to assign an affinity to this binding (7), there is general agreement that TeNT shows the highest affinity for GT1b and GD1b polysialogangliosides, although not nearly as ...

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“…Figure 2 illustrates the paroxysmal activity produced by tetanus toxin. Previous characterization of the action of tetanus toxin on spinal cord neurons in culture (Bergey et al, 1983(Bergey et al, , 1987 has demonstrated that following the addition of toxin there is a dose dependent latent period with no observable change seen with intracellular recordings of spontaneous electrical 11 activity. Presumably during this latent period the toxin is binding to the neuronal membrane and undergoing a subsequent membrane interaction or internalization.…”

Section: Effects Of Antibodies On Dindiug Of Tetanus Toxinmentioning

confidence: 99%

“…2, 3). All spinal cord neurons that are destined to develop tetanus-PDE will do so by 45-90 minutes after the addition of 100 ng/ml at 33-34 C. Because of the abundant synaptic connections in the spinal core cultures, 90-95% of the spinal cord neurons will show PDE after the addition of toxin (Bergey et al, 1983).…”

Section: Effects Of Antibodies On Convulsant Activity Produced By Tetmentioning

confidence: 99%

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Mechanisms of Action of Clostridial Neurotoxins on Dissociated Mouse Spinal Cord Neurons in Cell Culture

Bergey¹

1991

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Tetanus toxin: convulsant action on mouse spinal cord neurons in culture

Cited by 66 publications

References 52 publications

Intrinsic Dynamics in Neuronal Networks. II. Experiment

Intrinsic Dynamics in Neuronal Networks. II. Experiment

Neuronal Sensitivity to Tetanus Toxin Requires Gangliosides

Mechanisms of Action of Clostridial Neurotoxins on Dissociated Mouse Spinal Cord Neurons in Cell Culture

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