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. 2019 Jul 25;13(2):405–418. doi: 10.1016/j.stemcr.2019.06.005

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© 2019 The Authors

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

PMC Copyright notice

Planarian Stem Cells Display Higher Median Telomere Fluorescence Intensity than Immediate, Early, and Late Post-mitotic Descendant Cells

(A) Maximum confocal projections of representative planarian paraffin sections. Magenta box indicates the region shown below. Anterior is to the left, posterior is to the right, and dorsal is up.

(B) Telomere intensity maps and stacked bar graphs for the representative images in (A) show that stem cells have the longest telomeres when compared with post-mitotic progeny. The intensity maps display the nuclei colored according to their telomere fluorescence intensity. The stacked bar graphs represent the proportion of nuclei within a given category of intensity. smedwi-1 is chosen as the reference marker and set up to allocate in each range of intensity approximately one-fourth of the total cells. Magenta box indicates the region shown below and corresponds to the high magnifications in (A).

(C) The column scatterplot shows all the pooled nuclei from the total of planarians and cells indicated; the median from smedwi-1⁺ cells is higher than that of prog-1⁺ cells (two-tailed Mann-Whitney U test; p < 0.0001), whereas the median from prog-1⁺ cells is higher than that of agat-1⁺ cells (two-tailed Mann-Whitney U test; p < 0.0001). The median of smedwi-1⁺ cells is the highest when compared with the other conditions (Kruskal-Wallis test; p < 0.0001).

(D) The schematic represents the process of stem cell differentiation. The images show a representative subregion from one of the zones used for quantification. The telomere intensity map displays all the nuclei shown in the representative image colored according to their telomere fluorescence intensity. The population of smedwi-1⁺/SMEDWI-1⁺ has an increased number of stem cells with strong telomere intensity (either orange or red categories) when compared with its immediate progeny.

(E) The graph shows all the nuclei pooled from all planarians analyzed (four zones per planarian). The median telomere intensity is lower in the SMEDWI-1⁺/smedwi-1⁻ when compared with SMEDWI-1⁺/smedwi-1⁺ (two-tailed Mann-Whitney U test; p < 0.0001).

Arbitrary units are not comparable between (C) and (E) for being two independent experiments. tel, telomeres; n, number of planarians analyzed; a.u., arbitrary units of fluorescence. Scale bars, 1 mm (A and B, main images), 220 μm (A and B, high-magnification images), and 60 μm (D).

See also Figure S2.