Journal: Frontiers in Chemistry

Article Title: Bile Acids Gate Dopamine Transporter Mediated Currents

doi: 10.3389/fchem.2021.753990

Figure Lengend Snippet: OCA generates an electrical current through the DAT. (A) Schematic representation of oocyte collection, cRNA synthesis and injection, and TEVC technique. (B) Representative traces of currents recorded by TEVC (Vh = −60 mV) from oocytes expressing mDAT, without or with hTGR5, uninjected, or expressing hTGR5 alone. The oocytes were perfused with 30 µM DA or 10 µM OCA. (C) Mean of the maximal DA-associated and OCA-induced currents (I nA ± SE of 14–47 oocytes, 6–11 batches) in oocytes expressing mDAT (C, left) or mDAT plus TGR5 (C, right) . (D) Mean of the maximal OCA-induced currents in oocytes expressing the proteins indicated; one-way ANOVA F(2,74) = 36.13, p < 0.0001 followed by Tukey’s multiple comparison test (mDAT vs mDAT-hTGR5 p = 0.566, mDAT vs hTGR5 p < 0.0001). (E) Representative traces of currents recorded by TEVC (Vh = −60 mV) from oocytes tested with DA (30 µM) and OCA at the indicated concentrations. The current of each trace was normalized to the mean current of dopamine to reduce the variability between batches. (F) I/V relationships of DA (30 µM) and OCA (10 µM)-induced currents.

Article Snippet: Electrophysiological studies were performed using the Two-Electrode Voltage Clamp (TEVC) technique (Oocyte Clamp OC-725; Warner Instruments, Hamden, CT, United States).

Techniques: Injection, Expressing, Comparison

Journal: Frontiers in Chemistry

Article Title: Bile Acids Gate Dopamine Transporter Mediated Currents

doi: 10.3389/fchem.2021.753990

Figure Lengend Snippet: OCA modulates the mDAT-mediated leak current. (A) Representative current traces recorded with TEVC (Vh = −60 mV) in oocytes expressing mDAT perfused with 30 µM DA or 10 µM OCA in ND98 or Li98 buffer. (B) Mean of maximal currents under conditions shown in A (I nA ± SE of 6–28 oocytes, 4 batches. One-way ANOVA F(4,94) = 42.29, p < 0.0001 followed by Bonferroni’s multiple comparison test (DA vs OCA p > 0.999, DA vs DA in Li + p > 0.999, OCA vs OCA in Li + p = 0.0004, DA in Li + vs OCA in Li + p < 0.0001). (C) Representative trace from oocytes expressing mDAT perfused with 30 µM DA or 10 µM OCA in ND98 or TMA98 buffer. (D) Mean of maximal currents under conditions shown in C (I nA ± SE of 6–15 oocytes, 3 batches. One-way ANOVA F(4,51) = 86.94, p < 0.0001 followed by Bonferroni’s multiple comparison test (DA vs OCA p = 0.359, DA vs DA in TMA + p < 0.0001, OCA vs OCA in TMA + p = 0.059, DA in TMA + vs OCA in TMA + p > 0.999).

Article Snippet: Electrophysiological studies were performed using the Two-Electrode Voltage Clamp (TEVC) technique (Oocyte Clamp OC-725; Warner Instruments, Hamden, CT, United States).

Techniques: Expressing, Comparison

Journal: Frontiers in Chemistry

Article Title: Bile Acids Gate Dopamine Transporter Mediated Currents

doi: 10.3389/fchem.2021.753990

Figure Lengend Snippet: Intracellular calcium does not regulate OCA-induced currents. (A) Schematic representation of the EGTA injection technique. (B) Representative trace of current recorded by TEVC (Vh = −60 mV) in oocytes expressing mDAT and injected with 13 mM EGTA in intracellular solution 30 min before exposure to 30 µM DA or 10 µM OCA in ND98 buffer (top) and the mean of maximal transport-associated and OCA-induced currents (bottom) (I nA ± SE of 6–7 oocytes, 2 batches). (C) Mean of maximal OCA-induced currents in mDAT oocytes with or without injection of EGTA. Two-tailed Student’s t -test, p = 0.476.

Article Snippet: Electrophysiological studies were performed using the Two-Electrode Voltage Clamp (TEVC) technique (Oocyte Clamp OC-725; Warner Instruments, Hamden, CT, United States).

Techniques: Injection, Expressing, Two Tailed Test

Journal: Frontiers in Chemistry

Article Title: Bile Acids Gate Dopamine Transporter Mediated Currents

doi: 10.3389/fchem.2021.753990

Figure Lengend Snippet: LCA induces a current that parallels the OCA-induced current. (A) Structure of OCA and LCA (B) Representative trace of current recorded with TEVC (Vh = −60 mV) in oocytes expressing mDAT and perfused with 30 µM DA or 10 µM LCA in ND98 buffer (top) . Mean of maximal dopamine transport-associated and LCA-induced currents (I nA ± SE of 8 oocytes, 3 batches) (bottom) . (C) Mean of maximal currents elicited by OCA or LCA in mDAT-expressing oocytes ( n = 8; p = 0.22 by Student’s t -test).

Article Snippet: Electrophysiological studies were performed using the Two-Electrode Voltage Clamp (TEVC) technique (Oocyte Clamp OC-725; Warner Instruments, Hamden, CT, United States).

Techniques: Expressing