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Publication : A high-content platform for physiological profiling and unbiased classification of individual neurons.

First Author  DuBreuil DM Year  2021
Journal  Cell Rep Methods Volume  1
Issue  1 PubMed ID  34318289
Mgi Jnum  J:311671 Mgi Id  MGI:6729767
Doi  10.1016/j.crmeth.2021.100004 Citation  DuBreuil DM, et al. (2021) A high-content platform for physiological profiling and unbiased classification of individual neurons. Cell Rep Methods 1(1)
abstractText  High-throughput physiological assays lose single-cell resolution, precluding subtype-specific analyses of activation mechanism and drug effects. We demonstrate APPOINT (automated physiological phenotyping of individual neuronal types), a physiological assay platform combining calcium imaging, robotic liquid handling, and automated analysis to generate physiological activation profiles of single neurons at large scale. Using unbiased techniques, we quantify responses to sequential stimuli, enabling subgroup identification by physiology and probing of distinct mechanisms of neuronal activation within subgroups. Using APPOINT, we quantify primary sensory neuron activation by metabotropic receptor agonists and identify potential contributors to pain signaling. We expand the role of neuroimmune interactions by showing that human serum directly activates sensory neurons, elucidating a new potential pain mechanism. Finally, we apply APPOINT to develop a high-throughput, all-optical approach for quantification of activation threshold and pharmacologically validate contributions of ion channel families to optical activation.
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