title: Development of a spontaneously active dorsal root ganglia assay using multiwell multielectrode arrays.
abstract: In vitro phenotypic assays of sensory neuron activity are important tools for identifying potential analgesic compounds. These assays are typically characterized by hyperexcitable and/or abnormally, spontaneously active cells. Whereas manual electrophysiology experiments provide high-resolution biophysical data to characterize both in vitro models and potential therapeutic modalities (e.g., action potential characteristics, the role of specific ion channels, and receptors), these techniques are hampered by their low throughput. We have established a spontaneously active dorsal root ganglia (DRG) platform using multiwell multielectrode arrays (MEAs) that greatly increase the ability to evaluate the effects of multiple compounds and conditions on DRG excitability within the context of a cellular network. We show that spontaneous DRG firing can be attenuated with selective Na(+) and Ca(2+) channel blockers, as well as enhanced with K(+) channel blockers. In addition, spontaneous activity can be augmented with both the transient receptor potential cation channel subfamily V member 1 agonist capsaicin and the peptide bradykinin and completely blocked with neurokinin receptor antagonists. Finally, we validated the use of this assay by demonstrating that commonly used neuropathic pain therapeutics suppress DRG spontaneous activity. Overall, we have optimized primary rat DRG cells on a multiwell MEA platform to generate and characterize spontaneously active cultures that have the potential to be used as an in vitro phenotypic assay to evaluate potential therapeutics in rodent models of pain.
authors.affiliation: Genetically Defined Diseases, Bristol-Myers Squibb Company, Wallingford, Connecticut; and.
authors.forename: Kim
authors.initials: K
authors.lastname: Newberry
authors.affiliation: Genetically Defined Diseases, Bristol-Myers Squibb Company, Wallingford, Connecticut; and.
authors.forename: Shuya
authors.initials: S
authors.lastname: Wang
authors.affiliation: Genetically Defined Diseases, Bristol-Myers Squibb Company, Wallingford, Connecticut; and.
authors.forename: Nina
authors.initials: N
authors.lastname: Hoque
authors.affiliation: Leads Discovery and Optimization, Bristol-Myers Squibb Company, Wallingford, Connecticut.
authors.forename: Laszlo
authors.initials: L
authors.lastname: Kiss
authors.affiliation: Genetically Defined Diseases, Bristol-Myers Squibb Company, Wallingford, Connecticut; and.
authors.forename: Michael K
authors.initials: MK
authors.lastname: Ahlijanian
authors.affiliation: Genetically Defined Diseases, Bristol-Myers Squibb Company, Wallingford, Connecticut; and.
authors.forename: James
authors.initials: J
authors.lastname: Herrington
authors.affiliation: Genetically Defined Diseases, Bristol-Myers Squibb Company, Wallingford, Connecticut; and john.graef@bms.com.
authors.forename: John D
authors.initials: JD
authors.lastname: Graef
mesh_terms: D000200:Action Potentials / Q000187:drug effects / Q000502:physiology; D000818:Animals; D001920:Bradykinin / Q000494:pharmacology; D002121:Calcium Channel Blockers / Q000494:pharmacology; D002211:Capsaicin / Q000494:pharmacology; D002478:Cells, Cultured; D004622:Embryo, Mammalian; D005260:Female; D005727:Ganglia, Spinal / Q000166:cytology*; D006358:Hot Temperature; D049990:Membrane Transport Modulators / Q000494:pharmacology; D020748:Mibefradil / Q000494:pharmacology; D009474:Neurons / Q000187:drug effects / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D018689:Sensory System Agents / Q000494:pharmacology; D026941:Sodium Channel Blockers / Q000494:pharmacology; D012965:Sodium Chloride / Q000494:pharmacology; D013373:Substance P / Q000378:metabolism; D013779:Tetrodotoxin / Q000494:pharmacology
chemical_list: D002121:Calcium Channel Blockers; D049990:Membrane Transport Modulators; D018689:Sensory System Agents; D026941:Sodium Channel Blockers; D020748:Mibefradil; D013373:Substance P; D013779:Tetrodotoxin; D012965:Sodium Chloride; D002211:Capsaicin; D001920:Bradykinin
keywords: DRG; MEA; nociceptors; sensory; spontaneous activity