Suitability of Clonal NS26 Cells for Botulinum Neurotoxin Studies
Abstract
We examined the NS-26 neuroblastoma cell line for suitability as a model for BoNT investigation. These cells are derived from a sympathetic ganglion tumor and have been reported to synthesize, store and release ACh and to form cholinergic synapses with skeletal muscle. In the current study, NS-26 cells were found to express the SNARE proteins SNAP-25 and syntaxin and to release ACh in a voltage- and Ca2+ -dependent fashion. ACh release was assessed by the uptake and release of radiolabeled ACh or by the quenching of the fluorescent dye FM1-43. For qualitative measure of synaptic activity, the latter was found to be as reliable as the radioisotope method but was more rapid and required less sample manipulation. Exposure of NS-26 cells to 10 nM BoNT/A resulted in complete inhibition of evoked ACh release accompanied by cleavage of ~35% of SNAP-25. NS-26 cells were also sensitive to the inhibitory actions of BoNT/C1 and BoNT/E, exhibiting a similar potency ratio to that observed in mouse lethality assays. It is concluded that NS-26 can serve as a good model for BoNT research.
Document Details
- Document Type
- Technical Report
- Publication Date
- Aug 21, 2020
- Accession Number
- AD1108974
Entities
People
- Heather Manley
- Heidi Hoard-fruchey
- Michael Adler
Organizations
- United States Army Medical Research Institute of Chemical Defense