Sensitivity of embryonic rat dorsal root ganglia neurons to Clostridium botulinum neurotoxins

Clostridium botulinum neurotoxins (BoNT) are zinc dependent endopeptidases which, once internalised into the neuronal cytosol, block neurotransmission by proteolysis of membrane-associated proteins putatively involved in synaptic vesicle docking and fusion with the plasma membrane. Although many stu...

Full description

Saved in:
Bibliographic Details
Published in:Toxicon (Oxford) 2000-02, Vol.38 (2), p.245-258
Main Authors: Welch, Mary J., Purkiss, John R., Foster, Keith A.
Format: Article
Language:English
Subjects:
Animals
Bacteriology
Biological and medical sciences
Botulinum Toxins - toxicity
Calcium - physiology
Cells, Cultured
Clostridial neurotoxins
Clostridium botulinum
Culture
Dorsal root ganglion
endopeptidase
Fundamental and applied biological sciences. Psychology
Ganglia, Spinal - drug effects
Ganglia, Spinal - embryology
Microbiology
Pathogenicity, virulence, toxins, bacteriocins, pyrogens, host-bacteria relations, miscellaneous strains
Potassium - pharmacology
Rats
Rats, Sprague-Dawley
Secretion
SNAP-25
Substance P
Substance P - metabolism
Syntaxin
VAMP
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Clostridium botulinum neurotoxins (BoNT) are zinc dependent endopeptidases which, once internalised into the neuronal cytosol, block neurotransmission by proteolysis of membrane-associated proteins putatively involved in synaptic vesicle docking and fusion with the plasma membrane. Although many studies have used a variety of cellular systems to study the neurotoxins, most require relatively large amounts of toxin or permeabilisation to internalise the neurotoxin. We present here a primary culture of embryonic rat dorsal root ganglia (DRG) neurons that exhibits calcium-dependent substance P secretion when depolarised with elevated extracellular potassium and is naturally BoNT sensitive. The DRG neurons showed a different IC 50 for each of the toxins tested with a 1000 fold difference between the most and least potent neurotoxins (0.05, 0.3, 30 and ∼60 nM for A, C, F and B, respectively). BoNT/A cleavage of SNAP-25 was seen as early as 2 h, but substance P secretion was not significantly inhibited until 4 h intoxication and the effects of BoNT/A were observed for as long as 15 days. This primary neuronal culture system represents a new and sensitive cellular model for the in vitro study of the botulinum neurotoxins.
ISSN:0041-0101
1879-3150
DOI:10.1016/S0041-0101(99)00153-1