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. 2014 Jun 17;22(8):1530–1543. doi: 10.1038/mt.2014.86

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Copyright © 2014 American Society of Gene & Cell Therapy

This work is licensed under a Creative Commons Attribution 3.0 Unported License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/3.0/

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Human pluripotent stem cells (hPSC)-sensory express ASIC1/2 heteromers and ASIC3 homomeric ion channels. (a) Representative trace of an inward current in response to a pH drop from physiological (pH 7.4) to acidic (pH 6.0) conditions. (b) Concentration–response curve for hPSC-sensory expressed ASICs. Similarly to recombinantly expressed ASICs, the Hill slope is very steep (Hill slope = 3.9) with an IC₅₀ = pH 6.5 (n = 7). Inset: non-normalized peak current densities observed at pH 6.0 (22.2 ± 1.7 pA/pF, n = 7). (c) I_pH recovery from desensitization was measured by two pH 6.0 drops separated by a test recovery period. Currents were fully recovered with a recovery period between 120 and 240 seconds (τ_recovery = 111.1 ± 19.8 seconds). (d) Representative traces to pH6 before (black traces) and after subunit-selective blockers psalmotoxin (ASIC1a left panel), mambalgin-1 (ASIC1a/b 2a/b middle panel), APETx2 (ASIC3 right panel). (e) Quantification of toxin inhibition as shown in d. Psalmotoxin exhibited limited to no inhibition of I_pH in hPSC-sensory (0.9 ± 0.01 fraction of control, n = 11), whereas mambalgin-1 showed large inhibition demonstrating the presence of heteromeric ASICs (0.1 ± 0.01 fraction of control, n = 4). APETx2 showed small block (0.8 ± 0.1 fraction of control, n = 4) suggesting low levels of ASIC3 homomers.