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12/3/370    most recent
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This version was published on April 1, 2007
Journal of Biomolecular Screening, Vol. 12, No. 3, 370-377 (2007)
DOI: 10.1177/1087057106299163
© 2007 Society for Biomolecular Sciences

Primary Cultures of Embryonic Chicken Neurons for Sensitive Cell-Based Assay of Botulinum Neurotoxin: Implications for Therapeutic Discovery

Andrea M. Stahl

U.S. Army Medical Research Institute of Infectious Diseases, Frederick, Maryland

Gordon Ruthel

U.S. Army Medical Research Institute of Infectious Diseases, Frederick, Maryland, gordon.ruthel{at}amedd.army.mil

Edna Torres-Melendez

U.S. Army Medical Research Institute of Infectious Diseases, Frederick, Maryland

Tara A. Kenny

Target Structure-Based Drug Discovery Group, SAIC-Frederick Inc., NCI-Frederick, Frederick, Maryland

Rekha G. Panchal

Target Structure-Based Drug Discovery Group, SAIC-Frederick Inc., NCI-Frederick, Frederick, Maryland

Sina Bavari

U.S. Army Medical Research Institute of Infectious Diseases, Frederick, Maryland, sina.bavari{at}amedd.army.mil

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. (Journal of Biomolecular Screening 2007:370-377)

Key Words: botulinum neurotoxin • chick neurons • SNAP-25


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