1. ORL1 inhibits axon regeneration.
ORL1 inhibits axon regeneration. (A and B) Cortical neurons were transduced with lentiviral vector for control or ORL1 shRNA at 3 DIV. Axons were scraped at 8 DIV and allowed to regenerate for 3 days in the presence of vehicle or Nogo22 (100 nM). The microphotographs show ßIII-tubulin (in axons; green) and phalloidin (to stain F-actin; red) to illustrate the growth cones of cortical neurons in the middle of the scraped area (A). Scale bar, 100 μm. The graph shows quantification of axon regeneration (B). Means ± SE. n = 10 biological replicates, **P < 0.01, #not significant, one-way ANOVA followed by Tukey’s test. (C and D) Cortical neurons were transduced with lentiviral vector for control or ORL1 shRNA at 3 DIV. Axons were scraped at 8 DIV and allowed to regenerate for 3 days. Images are of the scraped areas from Ngr1+/+ and Ngr1−/− cultures (C). Scale bar, 100 μm. The graph shows quantification of axon regeneration (D). Means ± SE. n = 4 biological replicates, ***P < 0.005, *P < 0.05, Student’s two-tailed t test. (E and F) Cortical neurons were nucleofected with control vector or HA-ORL1. Axons were scraped at 8 DIV and allowed to regenerate for 3 days. Images are of the scraped area from Ngr1+/+ cultures (E). Scale bar, 100 μm. The graph shows quantification of axon regeneration (F). Data are means ± SE. n = 3 biological replicates, ***P < 0.005, Student’s two-tailed t test.
Yuichi Sekine et al., Sci. Signal. 2018;11:eaao4180
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