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Figure 1. 3 h treatment with Ibogaine leads to a long-lasting increase in GDNF expression that is mediated by the GDNF polypeptide. A) Cells were treated without (Con, lanes 1, 3, 5, and 7), or with (Ibo, lanes 2, 4, 6, and 8) 10 µM Ibogaine for 3 h. Cells were then extensively washed and incubated in fresh media for the indicated period. GDNF expression was analyzed by RT-polymerase chain reaction (RT-PCR). Histogram depicts the mean ratios of GDNF to Actin ± SD of 3 experiments. *P < 0.05; **P < 0.01, compared with control. B) Cells were treated without (Con), or with 10 µM Ibogaine for 3 h. Cells were then washed extensively and incubated in fresh media for the indicated period without (lower panel) or with (upper panel) 5 µg/ml actinomycin D (A/D). Histogram depicts the mean percentages of GDNF ± SD of 3 experiments. *P < 0.05; **P < 0.01, compared with 0 time. C) Cells were treated without (lanes 1 and 3) or with (lanes 2 and 4) 10 µM Ibogaine for 3 h. Cells were then extensively washed and incubated with fresh media in the absence (lanes 1 and 2) or presence (lanes 3 and 4) of 10 µg/ml anti-GDNF neutralizing antibodies for 3 h. Histogram depicts the mean ratios of GDNF to Actin ± SD of 3 experiments. **P < 0.01, lane 2 vs. 1, or lane 4 vs. 2.
