Primary cultures of embryonic chicken neurons for sensitive cell-based assay of botulinum neurotoxin: implications for therapeutic discovery.
Botulinum toxin is an exceedingly potent inhibitor of neurotransmission across the neuromuscular junction, causing flaccid paralysis and death. The potential for misuse of this deadly poison as a bioweapon has added a greater urgency to the search for effective therapeutics. The development of sensitive and efficient cell-based assays for the evaluation of toxin antagonists is crucial to the rapid and successful identification of therapeutic compounds. The authors evaluated the sensitivity of primary cultures from 4 distinct regions of the embryonic chick nervous system to botulinum neurotoxin A (BoNT/A) cleavage of synaptosomal-associated protein of 25 kD (SNAP-25). Although differences in sensitivity were apparent, SNAP-25 cleavage was detectable in neuronal cells from each of the 4 regions within 3 h at BoNT/A concentrations of 1 nM or lower. Co-incubation of chick neurons with BoNT/A and toxin-neutralizing antibodies inhibited SNAP-25 cleavage, demonstrating the utility of these cultures for the assay of BoNT/A antagonists.
Stahl
Andrea M
AM
U.S. Army Medical Research Institute of Infectious Diseases, Frederick, Maryland 21702, USA.
Ruthel
Gordon
G
Torres-Melendez
Edna
E
Kenny
Tara A
TA
Panchal
Rekha G
RG
Bavari
Sina
S
D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D001681:Biological Assay / Q000379:methods*; D019274:Botulinum Toxins, Type A / Q000037:antagonists & inhibitors* / Q000494:pharmacology*; D002478:Cells, Cultured; D002642:Chick Embryo; D004353:Drug Evaluation, Preclinical; D009474:Neurons / Q000166:cytology* / Q000187:drug effects*; D012680:Sensitivity and Specificity; D050825:Synaptosomal-Associated Protein 25 / Q000378:metabolism; D013997:Time Factors
D000906:Antibodies; D050825:Synaptosomal-Associated Protein 25; D019274:Botulinum Toxins, Type A