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A pharmacological model of TRPA1‐mediated nociception in zebrafish larvae for therapeutic discovery
Author(s) -
Ko Mee Jung,
Ganzen Logan C.,
Leung Yuk Fai,
Rijn Richard M.
Publication year - 2018
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2018.32.1_supplement.684.3
Subject(s) - zebrafish , transient receptor potential channel , agonist , in vivo , calcium imaging , pharmacology , calcium channel , antagonist , calcium , voltage dependent calcium channel , neuroscience , nociception , chemistry , receptor , medicine , biology , biochemistry , microbiology and biotechnology , gene
Nearly 50 million people in the United States currently suffer from chronic pain disorder (CPD). CPD accompanies unrelieved and persistent pain lasting longer than 3 months. For many patients, current medications inadequately treat their symptoms or come with significant adverse effects, suggesting the need to find novel treatments. One novel therapeutic target for CPD is a calcium channel called Transient Receptor Potential channel subfamily A1 (TRPA1). However, the lack of robust in vivo models to investigate TRPA1 has been perceived as a challenge to drug discovery. Zebrafish model has been utilized for discovering neuroactive compounds. Their increased locomotive/swimming behavior upon TRPA1 activation has been interpreted as a nociceptive‐like phenotype. To increase the robustness of this model, we employed a less volatile and less neurotoxic TRPA1 agonist. We validated TRPA1‐mediated responses by a selective TRPA1 antagonist. Here, we first characterized the basic pharmacology of mouse and zebrafish TRPA1 ligands in HEK293 cells transiently expressing TRPA1. We used FLIPR ® ‐based calcium assay to record calcium influx, which is a measure of TPPA1 function. Additionally, we measured swimming behavior of zebrafish larvae (5 days post‐fertilization) following TRPA1 activation using a ViewPoint Zebrabox. In our studies, treatment with 100 μM TRPA1 agonist (ASP7663) increased calcium influx in HEK293 cells and swimming behavior of zebrafish larvae. Pre‐incubation of TRPA1 antagonist (HC030031) ablated in vitro calcium influx in a dose‐dependent manner and TRPA1‐mediated locomotive behavior in zebrafish larvae. Our findings validate that the zebrafish larvae can act as an in vivo model to investigate TRPA1‐targeted drugs for CPD. The increased robustness of our model will allow for a successful target‐based drug screening. Support or Funding Information The current research has been funded by a Collaborative Research Award from the Purdue Institute of Integrative Neuroscience (PIIN). This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .