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Figure 6 | Frontiers in Zoology

Figure 6

From: Xenopus embryonic epidermis as a mucociliary cellular ecosystem to assess the effect of sex hormones in a non-reproductive context

Figure 6

Effects of sex steroids on the MCE cellular ecosystem. A,B) Effects of estradiol (E2), testosterone (T) and ethynyl-E2 (EE2) on the total cell abundance in MCE, based on DAPI nuclei staining (A), and on the total amount of proliferative cells (pH3 positive/field) (B). Analyses by confocal microscopy in embryos at st 30, d.f. = 21. C) Proliferative cells in E2 treated MCE from ectoderm explants. IHC for pH3 and DAPI. Confocal microscopy, 2 μm stack. Data representative of 18 samples/condition. D,E) Changes in cell population enrichment mediated by sex steroids in Xenopus epidermal MCE. Data based on the ratio of cell counting of each population over the total number of cells at st 30 (DAPI) (D) or st 40 (SEM micrographs) (E). Data are representative of 48 (st 30, Confocal imaging of molecular makers) or 24 (st 40) embryos by sex steroid treatment. +/++ concentrations = 1x10-7 M / 1x10-5 M. MC = multiciliated, vMR or rMR = vesicle or ridged mitochondrion-rich, MS = mucus-secreting cells. A,B,D,E) Baseline established for control group = 0. Mean ± SD, “a” = control group; Letter code: a indicates no difference with control, all other letters indicate significant difference with the control, shared letters indicate no significant differences between the corresponding conditions. One-way ANOVA, Tukey comparison test, p < 0.05. F) Disruption of cell population arrangement caused by EE2 in embryos at st 30. We observed an increase of MC (tuba1a-b, green) and MR (atp6v1a, red) cells, and the presence of adjacent MC cells. Double FISH and DAPI. Confocal microscopy, 2 μm stack, 24 embryos/condition. Right panels: Increase of the number of MC cells and disrupted epithelial architecture caused by EE2 SEM, st 40, 16 embryos/condition. G) Perturbed epidermal MCE arrangement and morphology provoked by T. SEM, st 40, 16 embryos/condition.

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