A high content imaging assay for identification of Botulinum neurotoxin inhibitors.

Date

2014-11-14

Author

Kota, KP
Soloveva, V
Wanner, LM
Gomba, G
Kiriş, Erkan
Panchal, RG
Kane, CD
Bavari, S

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Synaptosomal-associated protein-25 (SNAP-25) is a component of the soluble NSF attachment protein receptor (SNARE) complex that is essential for synaptic neurotransmitter release. Botulinum neurotoxin serotype A (BoNT/A) is a zinc metalloprotease that blocks exocytosis of neurotransmitter by cleaving the SNAP-25 component of the SNARE complex. Currently there are no licensed medicines to treat BoNT/A poisoning after internalization of the toxin by motor neurons. The development of effective therapeutic measures to counter BoNT/A intoxication has been limited, due in part to the lack of robust high-throughput assays for screening small molecule libraries. Here we describe a high content imaging (HCI) assay with utility for identification of BoNT/A inhibitors. Initial optimization efforts focused on improving the reproducibility of inter-plate results across multiple, independent experiments. Automation of immunostaining, image acquisition, and image analysis were found to increase assay co

Subject Keywords

Neurotoxicity, High content imaging system, Clostridium botulinum, Botulinum neurotoxin, Neurobiology, Neuroscience, Issue 93, Neuroscience

URI

https://hdl.handle.net/11511/37123

Journal

Journal of visualized experiments : JoVE

DOI

https://doi.org/10.3791/51915

Collections

Department of Biology, Article

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Citation Formats

K. Kota et al., “A high content imaging assay for identification of Botulinum neurotoxin inhibitors.,” Journal of visualized experiments : JoVE, 2014, Accessed: 00, 2020. [Online]. Available: https://hdl.handle.net/11511/37123.