Figure 1
From: Light-controlled inhibition of malignant glioma by opsin gene transfer
ChETA gene transfer and electrophysiological recordings of human glioma cells. (a) Schematic drawings of the CMV-ChETA-eYFP and cfos-ChETA-eYFP lentivirus vector constructs. (b) Positive ChETA expression (green) was observed in normal neurons, astrocytes, and U87/H4 glioma cell lines transfected with CMV-ChETA-eYFP lentivirus. Bar=20 μm. (c) Schematic drawing of the arrangement of the lamps in the blue LED constructed for the light illumination; the average light intensity was 1.64 mW/mm2. (d) The constructed blue LED used to illuminate the cells in six-well plates. (e) Quantification of the percentage of viable neurons, astrocytes, and U87/H4 cells in the control and ChETA+light groups. (f) Positive ChETA expression was seen in the U87 and H4 glioma cells but not in rat astrocytes (As) transfected with cfos-ChETA-eYFP lentivirus; bar=50 μm. Top panel: bright field; bottom panel: ChETA expression (shown in green). (g) Whole-cell patch clamping was used to record light-evoked photocurrents from ChETA-expressing U87 cells. (h) Light stimulation (500 ms) induced depolarization and a recovery current. (i) The peak and steady-state amplitude of the induced depolarization current, n=12. (j) A ChETA channel current spike was induced by 1-, 21-, 51-, 101-, and 201-ms-light pulses. (k) Spike trains induced by paired light stimulations with 25-, 50-, 75-, 100-, and 200-ms intervals. (l) Spike trains were induced before (top panel) and after 1 h (bottom panel) of light illumination (10 Hz). **P<0.01
