Xenbase Image ID: 146117

Figure 6. Lack of retrograde opsin trafficking in mature rod OSs. A: Retinas were dissected and imaged for green and red fluorescence 2 days or 6 days after photoconversion at 21–22 dpf, or without photoconversion at 21–22 dpf. The intensity of green and red fluorescence was measured in ImageJ and the percentage of Opsin-Dend2 protein along the longitudinal axis of the OS was calculated by normalizing to the intensities at the crosspoint (100%). For OSs 2 days and 6 days after photoconversion, old Opsin-Dend2 protein concentrations in the disks from the crosspoint toward the base of the OS were normalized to their concentrations at the crosspoint. Likewise for nonphotoconverted OSs (0 d), the concentration distribution of Opsin-Dend2 around the bottom of the OS was normalized to the intensity closest to 50% of the maximum concentration along the OS axis. There were no significant differences among 0 day, 2 days, and 6 days in terms of the distribution of Opsin-Dend2. Thus, there was no retrograde trafficking in 21–22 dpf rod photoreceptors. For 0 and 2 days after photoconversion, n = 20 rod photoreceptors from three tadpoles were used for analysis. For 6 days after photoconversion, n = 10 rod photoreceptors from three tadpoles were used for analysis. The data points are represented by mean ± SE. B: The fluorescence intensities of Opsin-Dend2 in the rod OSs from the tadpoles at 7 (blue) and 21–22 (magenta) dpf without photoconversion were normalized to the highest intensities along the axes, and plotted along the longitudinal axis of the OS. C: The central regions (1.3 μm width) of OSs, as shown in B (light green), were used to calculate the standard deviations of normalized fluorescence intensities. Concentrations of Opsin-Dend2 along the longitudinal axis of rod photoreceptors were less variable in 7 dpf than in 21–22 dpf tadpoles. For both 7 and 21–22 dpf, n = 20 rod photoreceptors from three tadpoles were used for the calculations. ***P < 0.001 by the Mann–Whitney Rank Sum Test. Error bars represent standard deviation in C. Image published in: Tian G et al. (2014) Copyright © 2014. Image reproduced with permission of the Publisher, John Wiley & Sons. Larger Image Printer Friendly View

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