title: Primary cultures of embryonic chicken neurons for sensitive cell-based assay of botulinum neurotoxin: implications for therapeutic discovery.
abstract: 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.
authors.affiliation: U.S. Army Medical Research Institute of Infectious Diseases, Frederick, Maryland 21702, USA.
authors.forename: Andrea M
authors.initials: AM
authors.lastname: Stahl
authors.forename: Gordon
authors.initials: G
authors.lastname: Ruthel
authors.forename: Edna
authors.initials: E
authors.lastname: Torres-Melendez
authors.forename: Tara A
authors.initials: TA
authors.lastname: Kenny
authors.forename: Rekha G
authors.initials: RG
authors.lastname: Panchal
authors.forename: Sina
authors.initials: S
authors.lastname: Bavari
mesh_terms: 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
chemical_list: D000906:Antibodies; D050825:Synaptosomal-Associated Protein 25; D019274:Botulinum Toxins, Type A