XB-IMG-159732
Xenbase Image ID: 159732
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Fig. 3. Quantitative analyses of intracellular Ca2+ activities in the neural plate. (A) Representative results of quantitative analyses of a Ca2+ fluctuation in an
untreated (left), 2APB-treated (middle) and nifedipine-treated (right) embryo. R-GECO1.0 and EGFP were introduced into the left side of the NPs. (Top) EGFP
fluorescence images at the start of time-lapse recording; (middle) time projections of Ca2+ activity (R-GECO1.0/EGFP); and (bottom) time projections of
reconstructed circles corresponding to the area of the Ca2+ transient, extracted by image processing. Solid yellow lines indicate the midline; dotted lines indicate the
border between neural and non-neural tissues. The reconstructed circles were color-coded based on the quartiles of the area distributions in an untreated embryo.
(B) The mean number of Ca2+ transients during NTC. The time interval of observations was 20 s. The data are shown as smoothed curves, obtained by averaging
the originally counted numbers at a given time point with those of neighboring time points (50 time points total). The shaded area indicates the s.d. (C) The number of
single-cell Ca2+ transients in the early and late phases of NTC. **P<0.01 and ***P<0.001, two-sided Mann–Whitney U-test. (D) The number of multicellular Ca2+
transients in the early and late phases of NTC. **P<0.01, two-sided Mann–Whitney U-test. (E) The ratio of the total area of the multicellular or single-cell Ca2+
transients in the early and late phases of NTC in untreated embryos. ***P<0.001, two-sidedWilcoxon signed rank test. (F) Representative data for the closing speed
(black) and the total R-GECO1.0 intensity values at the tissue level (red) during NTC. The closing speed is shown as a smoothed curve, obtained by averaging the
rate determined from image analyses (gray) at a given time point with those of neighboring time points (ten time points total). Overlaps between temporary increases
in closing speed and intense Ca2+ transients are highlighted (yellow). (G) Mean cross-correlation coefficients between the closing speed and the total R-GECO1.0
value. The shaded area indicates s.d. n=19 (untreated), 5 (2APB-treated) and 6 (nifedipine-treated) embryos. Results in B-E andGare for the same data set. In C-E,
the box and whiskers of these plots indicate maximum, third quarter, median, first quarter and minimum values of each group, respectively, from upper side to the
bottom of the graphs. Data beyond the end of the whiskers, the upper limit of whiskers is 1.5 times the box length, are shown as outliers. Image published in: Suzuki M et al. (2017) Copyright © 2017. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License. Larger Image Printer Friendly View |