Author(s): Coyne L, Shan M, Przyborski SA, Hirakawa R, Halliwell RF
Human pluripotent stem cells have enormous potential value in neuropharmacology and drug discovery yet there is little data on the major classes and properties of receptors and ion channels expressed by neurons derived from these stem cells. Recent studies in this lab have therefore used conventional patch-clamp electrophysiology to investigate the pharmacological properties of the ligand and voltage-gated ion channels in neurons derived and maintained in vitro from the human stem cell (hSC) line, TERA2.cl.SP12.
TERA2.cl.SP12 stem cells were differentiated with retinoic acid and used in electrophysiological experiments 28–50 days after beginning differentiation. HSC-derived neurons generated large whole cell currents with depolarizing voltage steps (−80 to 30 mV) comprised of an inward, rapidly inactivating component and a delayed, slowly deactivating outward component. The fast inward current was blocked by the sodium channel blocker tetrodotoxin (0.1 μM) and the outward currents were significantly reduced by tetraethylammonium ions (TEA, 5 mM) consistent with the presence of functional Na and K ion channels. Application of the inhibitory neurotransmitters, GABA (0.1–1000 μM) or glycine (0.1–1000 μM) evoked concentration dependent currents. The GABA currents were inhibited by the convulsants, picrotoxin (10 μM) and bicuculline (3 μM), potentiated by the NSAID mefenamic acid (10–100 μM), the general anaesthetic pentobarbital (100 μM), the neurosteroid allopregnanolone and the anxiolytics chlordiazepoxide (10 μM) and diazepam (10 μM) all consistent with the expression of GABAA receptors. Responses to glycine were reversibly blocked by strychnine (10 μM) consistent with glycine-gated chloride channels. The excitatory agonists, glutamate (1–1000 μM) and NMDA (1–1000 μM) activated concentration-dependent responses from hSC-derived neurons. Glutamate currents were inhibited by kynurenic acid (1 mM) and NMDA responses were blocked by MgCl2 (2 mM) in a highly voltage-dependent manner.
Together, these findings show that neurons derived from human stem cells develop an array of functional receptors and ion channels with a pharmacological profile in keeping with that described for native neurons. This study therefore provides support for the hypothesis that stem cells may provide a powerful source of human neurons for future neuropharmacological studies.
Author(s): Colborn T
Author(s): Herbert MR
Author(s): Rauh VA, Garfinkel R, Perera FP, Andrews HF, Hoepner L, et al.
Author(s): Grandjean P, Landrigan PJ
Author(s): Grandjean P, Landrigan PJ
Author(s): Makris SL, Raffaele K, Allen S, Bowers WJ, Hasset U
Author(s): Middaugh LD, Dow-Edwards D, Li AA
Author(s): Cooper RL, Lamb JC, Barlow SM
Author(s): Doe JE, Boobis AR, Blacker A, Dellarco V, Doerrer NG, et al.
Author(s): Couillard-Despres S, Quehl E, Altendorfer K, Bentley K, Brady AM, et al.
Author(s): Hill EJ, Woehrling EK, Prince M, Coleman MD
Author(s): Hou Z, Zhang J, Schwartz MP, Stewart R, Page CD, et al.
Author(s): Laurenza I, Pallocca G, Mennecozzi M, Scelfo B, Pamies D, et al.
Author(s): Menzner AK, AbolpourMofrad S, Friedrich O, Gilbert DF
Author(s): Pallocca G, Fabbri M, Sacco MG, Gribaldo L, Pamies D, et al.
Author(s): Haile Y, Fu W, Shi B, Westaway D, Baker G, et al.
Author(s): Andrews PW
Author(s): Kuenzel K, Friedrich O, Gilbert DF
Author(s): Pleasure SJ, Lee VM
Author(s): Pleasure SJ, Page C, Lee VM
Author(s): Stewart R, Coyne L, Lako M, Halliwell RF, Przyborski SA
Author(s): Hsu TC, Liu KK, Chang HC, Hwang E, Chao JI
Author(s): Megiorni F, Mora B, Indovina P, Mazzilli MC
Author(s): Paquet-Durand F, Tan S, Bicker G
Author(s): Podrygajlo G, Tegenge MA, Gierse A
Author(s): Yao ZX, Han Z, Xu J, Greeson J, Lecanu L, et al.
Author(s): AbolpourMofrad S, Kuenzel K, Friedrich O, Gilbert DF