Persistence of Botulinum Neurotoxin a Subtypes 1-5 in Primary Rat Spinal Cord Cells

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2014 Mar 4

PMID 24587301

Citations 52

Authors

Regina Clare Meyer Whitemarsh

William Howard Tepp

Eric Arthur Johnson

Sabine Pellett

Affiliations

Soon will be listed here.

Abstract

Botulinum neurotoxins (BoNTs) are the most poisonous substances known and cause the severe disease botulism. BoNTs have also been remarkably effective as therapeutics in treating many neuronal and neuromuscular disorders. One of the hallmarks of BoNTs, particularly serotype A, is its long persistence of 2-6 months in patients at concentrations as low as fM or pM. The mechanisms for this persistence are currently unclear. In this study we determined the persistence of the BoNT/A subtypes 1 through 5 in primary rat spinal neurons. Remarkably, the duration of intracellular enzymatic activity of BoNT/A1, /A2, /A4 and /A5 was shown to be at least 10 months. Conversely, the effects of BoNT/A3 were observed for up to ∼5 months. An intermittent dosing with BoNT/E showed intracellular activity of the shorter acting BoNT/E for 2-3 weeks, followed by reoccurrence and persistence of BoNT/A-induced SNAP-25 cleavage products.

Citing Articles

Cryo-EM structure of the botulinum neurotoxin A/SV2B complex and its implications for translocation.

Khanppnavar B, Leka O, Pal S, Korkhov V, Kammerer R Nat Commun. 2025; 16(1):1224.

PMID: 39934119 PMC: 11814414. DOI: 10.1038/s41467-025-56304-z.

Potency Evaluations of Recombinant Botulinum Neurotoxin A1 Mutants Designed to Reduce Toxicity.

Viravathana P, Tepp W, Bradshaw M, Przedpelski A, Barbieri J, Pellett S Int J Mol Sci. 2024; 25(16).

PMID: 39201641 PMC: 11355004. DOI: 10.3390/ijms25168955.

The Degradation of Botulinum Neurotoxin Light Chains Using PROTACs.

Tsai Y, Kozar L, Mawi Z, Ichtchenko K, Shoemaker C, McNutt P Int J Mol Sci. 2024; 25(13).

PMID: 39000579 PMC: 11242356. DOI: 10.3390/ijms25137472.

Embracing the Versatility of Botulinum Neurotoxins in Conventional and New Therapeutic Applications.

Rasetti-Escargueil C, Palea S Toxins (Basel). 2024; 16(6).

PMID: 38922155 PMC: 11209287. DOI: 10.3390/toxins16060261.

A DARPin promotes faster onset of botulinum neurotoxin A1 action.

Leka O, Wu Y, Zanetti G, Furler S, Reinberg T, Marinho J Nat Commun. 2023; 14(1):8317.

PMID: 38110403 PMC: 10728214. DOI: 10.1038/s41467-023-44102-4.

References

Whitemarsh R, Tepp W, Bradshaw M, Lin G, Pier C, Scherf J . Characterization of botulinum neurotoxin A subtypes 1 through 5 by investigation of activities in mice, in neuronal cell cultures, and in vitro. Infect Immun. 2013; 81(10):3894-902. PMC: 3811745. DOI: 10.1128/IAI.00536-13. View

Heap J, Pennington O, Cartman S, Minton N . A modular system for Clostridium shuttle plasmids. J Microbiol Methods. 2009; 78(1):79-85. DOI: 10.1016/j.mimet.2009.05.004. View

Chakraborty S, Nazmi A, Dutta K, Basu A . Neurons under viral attack: victims or warriors?. Neurochem Int. 2010; 56(6-7):727-35. PMC: 7115389. DOI: 10.1016/j.neuint.2010.02.016. View

Fernandez-Salas E, Steward L, Ho H, Garay P, Sun S, Gilmore M . Plasma membrane localization signals in the light chain of botulinum neurotoxin. Proc Natl Acad Sci U S A. 2004; 101(9):3208-13. PMC: 365768. DOI: 10.1073/pnas.0400229101. View

Gimenez D, Gimenez J . The typing of botulinal neurotoxins. Int J Food Microbiol. 1995; 27(1):1-9. DOI: 10.1016/0168-1605(94)00144-u. View

Bradshaw M, Marshall K, Heap J, Tepp W, Minton N, Johnson E . Construction of a nontoxigenic Clostridium botulinum strain for food challenge studies. Appl Environ Microbiol. 2009; 76(2):387-93. PMC: 2805223. DOI: 10.1128/AEM.02005-09. View

SCHANTZ E, Johnson E . Properties and use of botulinum toxin and other microbial neurotoxins in medicine. Microbiol Rev. 1992; 56(1):80-99. PMC: 372855. DOI: 10.1128/mr.56.1.80-99.1992. View

Adler M, Keller J, Sheridan R, Deshpande S . Persistence of botulinum neurotoxin A demonstrated by sequential administration of serotypes A and E in rat EDL muscle. Toxicon. 2000; 39(2-3):233-43. DOI: 10.1016/s0041-0101(00)00120-3. View

Foran P, Mohammed N, Lisk G, Nagwaney S, Lawrence G, Johnson E . Evaluation of the therapeutic usefulness of botulinum neurotoxin B, C1, E, and F compared with the long lasting type A. Basis for distinct durations of inhibition of exocytosis in central neurons. J Biol Chem. 2002; 278(2):1363-71. DOI: 10.1074/jbc.M209821200. View

10.

Dineen S, Bradshaw M, Johnson E . Neurotoxin gene clusters in Clostridium botulinum type A strains: sequence comparison and evolutionary implications. Curr Microbiol. 2003; 46(5):345-52. DOI: 10.1007/s00284-002-3851-1. View

11.

Eleopra R, Tugnoli V, Rossetto O, De Grandis D, Montecucco C . Different time courses of recovery after poisoning with botulinum neurotoxin serotypes A and E in humans. Neurosci Lett. 1998; 256(3):135-8. DOI: 10.1016/s0304-3940(98)00775-7. View

12.

Dover N, Barash J, Hill K, Xie G, Arnon S . Molecular characterization of a novel botulinum neurotoxin type H gene. J Infect Dis. 2013; 209(2):192-202. DOI: 10.1093/infdis/jit450. View

13.

Pier C, Chen C, Tepp W, Lin G, Janda K, Barbieri J . Botulinum neurotoxin subtype A2 enters neuronal cells faster than subtype A1. FEBS Lett. 2010; 585(1):199-206. PMC: 3041152. DOI: 10.1016/j.febslet.2010.11.045. View

14.

Barash J, Arnon S . A novel strain of Clostridium botulinum that produces type B and type H botulinum toxins. J Infect Dis. 2013; 209(2):183-91. DOI: 10.1093/infdis/jit449. View

15.

Keller J, Neale E, Oyler G, Adler M . Persistence of botulinum neurotoxin action in cultured spinal cord cells. FEBS Lett. 1999; 456(1):137-42. DOI: 10.1016/s0014-5793(99)00948-5. View

16.

Dover N, Barash J, Arnon S . Novel Clostridium botulinum toxin gene arrangement with subtype A5 and partial subtype B3 botulinum neurotoxin genes. J Clin Microbiol. 2009; 47(7):2349-50. PMC: 2708499. DOI: 10.1128/JCM.00799-09. View

17.

Hill K, Smith T, Helma C, Ticknor L, Foley B, Svensson R . Genetic diversity among Botulinum Neurotoxin-producing clostridial strains. J Bacteriol. 2006; 189(3):818-32. PMC: 1797315. DOI: 10.1128/JB.01180-06. View

18.

de Paiva A, Meunier F, Molgo J, Aoki K, Dolly J . Functional repair of motor endplates after botulinum neurotoxin type A poisoning: biphasic switch of synaptic activity between nerve sprouts and their parent terminals. Proc Natl Acad Sci U S A. 1999; 96(6):3200-5. PMC: 15919. DOI: 10.1073/pnas.96.6.3200. View

19.

Luquez C, Raphael B, Maslanka S . Neurotoxin gene clusters in Clostridium botulinum type Ab strains. Appl Environ Microbiol. 2009; 75(19):6094-101. PMC: 2753052. DOI: 10.1128/AEM.01009-09. View

20.

Fernandez-Salas E, Ho H, Garay P, Steward L, Aoki K . Is the light chain subcellular localization an important factor in botulinum toxin duration of action?. Mov Disord. 2004; 19 Suppl 8:S23-34. DOI: 10.1002/mds.20006. View