Acute botulinum-like intoxication by tetanus neurotoxin in mice

Matsuda, Morihiro; Sugimoto, Nakaba; Ozutsumi, Kunihiro; Hirai, Toshihiro

doi:10.1016/0006-291x(82)90708-2

Cited by 42 publications

(23 citation statements)

References 15 publications

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“…Lipid monolayer studies have clearly documented the ability of 10 À8 M TeNT to interact with acidic lipids (Schiavo et al 1990a). Similar concentrations are routinely used with cells in culture, with in vivo injections into the hippocampus (Mellanby et al 1984) or in experiments of £accid paralysis in mice treated with one thousand times the mouse LD50 (Matsuda et al 1982). On the other hand, in clinical tetanus and botulism, the concentrations of TeNT and BoNTs at the periphery are sub-picomolar.…”

Section: Neurospecific Bindingmentioning

confidence: 99%

Tetanus and botulinum neurotoxins: mechanism of action and therapeutic uses

Pellizzari

Rossetto

Schiavo

et al. 1999

Phil. Trans. R. Soc. Lond. B

261

197

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The clostridial neurotoxins responsible for tetanus and botulism are proteins consisting of three domains endowed with di¡erent functions: neurospeci¢c binding, membrane translocation and proteolysis for speci¢c components of the neuroexocytosis apparatus. Tetanus neurotoxin (TeNT) binds to the presynaptic membrane of the neuromuscular junction, is internalized and transported retroaxonally to the spinal cord. The spastic paralysis induced by the toxin is due to the blockade of neurotransmitter release from spinal inhibitory interneurons. In contrast, the seven serotypes of botulinum neurotoxins (BoNTs) act at the periphery by inducing a £accid paralysis due to the inhibition of acetylcholine release at the neuromuscular junction. TeNT and BoNT serotypes B, D, F and G cleave speci¢cally at single but di¡erent peptide bonds, of the vesicle associated membrane protein (VAMP) synaptobrevin, a membrane protein of small synaptic vesicles (SSVs). BoNT types A, C and E cleave SNAP-25 at di¡erent sites located within the carboxyl-terminus, while BoNT type C additionally cleaves syntaxin. The remarkable speci¢city of BoNTs is exploited in the treatment of human diseases characterized by a hyperfunction of cholinergic terminals.

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Section: Neurospecific Bindingmentioning

confidence: 99%

Tetanus and botulinum neurotoxins: mechanism of action and therapeutic uses

Pellizzari

Rossetto

Schiavo

et al. 1999

Phil. Trans. R. Soc. Lond. B

261

197

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“…The dual uptake mechanism described here for BoNT/E parallels to some extent the more complex trafficking of tetanus toxin that also seems to enter peripheral nerves via both low-and highaffinity pathways. At low doses, tetanus toxin is mainly directed away from motor terminals (Lalli et al, 2003), but it is released locally if large quantities are presented; this phenomenon causes a switch from spastic to flaccid paralysis in severe cases of tetanus intoxication (Matsuda et al, 1982).…”

Section: Botulinum Toxin E Exploits Two Internalization Routes 415mentioning

confidence: 99%

Two Protein Trafficking Processes at Motor Nerve Endings Unveiled by Botulinum Neurotoxin E

Lawrence¹,

Wang²,

Chion³

et al. 2007

The Journal of Pharmacology and Experimental Therapeutics

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The unique ability of a family of botulinum neurotoxins to block neuroexocytosis specifically-by selective interaction with peripheral cholinergic nerve endings, endocytotic uptake, translocation to the cytosol, and enzymic cleavage of essential proteins-underlies their increasing therapeutic applications. Although clinical use of type A is most widespread due to its prolonged inactivation of the synaptosomal-associated protein of 25 kDa, botulinum neurotoxin E cleaves this same target but at a different bond and exhibits faster onset of neuromuscular paralysis. Herein, insights were gained into the different dynamics of action of types A and E toxins, which could help in designing variants with new pharmacological profiles. Natural and recombinant type E dichain forms showed similar proteolytic and neuromuscular paralytic activities. The neuroparalysis induced by type E toxin was accelerated between 21 and 35°C and attenuated by bafilomycin A1. Temperature elevation also revealed an unanticipated bipartite dose response indicative of two distinct internalization processes, one being independent of temperature and the other dependent. Although elevating the temperature also hastened intoxication by type A, a second uptake mechanism was not evident. Increasing the frequency of nerve stimulation raised the uptake of type E via both processes, but the enhanced trafficking through the temperaturedependent pathway was only seen at 35°C. These novel observations reveal that two membrane retrieval mechanisms are operative at motor nerve terminals which type E toxin exploits to gain entry via an acidification-dependent step, whereas A uses only one.Seven serotypes (termed A-G) of botulinum neurotoxin (BoNT), produced by the bacteria Clostridium botulinum, cause the disorder botulism that is characterized by flaccid neuromuscular paralysis (Dolly et al., 2002). BoNTs are proteins (mol. wt. 150 ϫ 10 3 ) with a heavy chain (ϳ100 ϫ 10 3 ) and light chain (50 ϫ 10 3 ) linked by a disulfide bond and noncovalent interactions. They are synthesized as singlechain (SC) precursors that require proteolytic nicking to yield the potent dichain (DC) forms. Each acts by inhibiting acetylcholine release via a complex multiphasic mechanism (Simpson, 1980(Simpson, , 2004: binding to high-affinity ectoacceptors on cholinergic nerve endings (Dolly et al., 1984), endocytotic uptake (Black and Dolly, 1986a,b), translocation of a toxic moiety, and specific cleavage of proteins essential for the release of transmitters from vesicles (Schiavo et al., 2000). Their heavy chain contributes to binding (Poulain et al., 1989) plus internalization (Dolly et al., 1994). Synaptic vesicle proteins have been identified as acceptors for some serotypes: SV2 for BoNT/A (Dong et al., 2006; Mahrhold et al., 2006) and synaptotagmins I and II for BoNT/B and BoNT/G (Nishiki et al., 1994(Nishiki et al., , 1996 Dong et al., 2003;Rummel et al., 2004a). An intact DC is needed for translocation (de Paiva et al., 1993) of the light chain to the presynaptic c...

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“…In this context, it is notable that motor nerve terminals appear to possess two distinct pools of TeTx acceptor. In addition to the aforementioned productive acceptor that underlies retrograde trafficking leading to the spastic paralysis typical of tetanus, low affinity binding sites exist that apparently release the neurotoxin proximal to its synaptic site of internalization; hence, at high doses TeTx induces flaccid paralysis at neuromuscular junctions in vitro (22) and in vivo (23). These various findings highlight that TeTx, devoid of toxicity, will provide the most effective vehicle for targeted delivery to the central nervous system, being functional at very low concentrations by targeting the high affinity acceptor, which enters the retrograde trafficking pathway and culminates in translocation of LC into the neuronal cytosol.…”

mentioning

confidence: 99%

Recombinant Forms of Tetanus Toxin Engineered for Examining and Exploiting Neuronal Trafficking Pathways

Yan¹,

Foran²,

Lawrence³

et al. 2001

Journal of Biological Chemistry

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Tetanus toxin is a fascinating, multifunctional protein that binds to peripheral neurons, undergoes retrograde transport and trans-synaptic transfer to central inhibitory neurons where it blocks transmitter release, thereby, causing spastic paralysis. As a pre-requisite for exploiting its unique trafficking properties, a novel recombinant single chain was expressed at a high level in Escherichia coli as a soluble, easily purifiable protein. It could be activated with enterokinase to produce a dichain that matched native toxin in terms of proteolytic and neuroinhibitory activities, as well as induction of spastic paralysis in mice. Importantly, nicking was not essential for protease activity. Substitution of Glu 234 by Ala created a protease-deficient atoxic form, which blocked the neuroparalytic action of tetanus toxin in vitro, with equal potency to its heavy chain; but, the mutant proved >30-fold more potent in preventing tetanus in mice. This observation unveils differences between the intoxication processes resulting from retrograde transport of toxin in vivo and its local uptake into peripheral or central nerves in vitro, dispelling a popularly held belief that the heavy chain is the sole determinant for efficient trafficking. Thus, this innocuous mutant may be a useful vehicle, superior to the heavy chain, for drug delivery to central neurons.

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Acute botulinum-like intoxication by tetanus neurotoxin in mice

Cited by 42 publications

References 15 publications

Tetanus and botulinum neurotoxins: mechanism of action and therapeutic uses

Tetanus and botulinum neurotoxins: mechanism of action and therapeutic uses

Two Protein Trafficking Processes at Motor Nerve Endings Unveiled by Botulinum Neurotoxin E

Recombinant Forms of Tetanus Toxin Engineered for Examining and Exploiting Neuronal Trafficking Pathways

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