Abstract
The Lugol’s staining method has been widely used to detect changes in the maintenance of stem cell fate in the columella root cap of Arabidopsis roots since the late 1990s. However, various limitations of this method demand for additional or complementary new approaches. For instance, it is unable to reveal the division rate of columella root cap stem cells. Here we report that, by labeling dividing stem cells with 5-ethynyl-2′-deoxyuridine (EdU), the number and distribution of their labeled progeny can be studied so that the division rate of stem cells can be measured quantitatively and in addition, that the progression of stem cell progeny differentiation can be assessed in combination with Lugol’s staining. EdU staining takes few hours and visualization of the stain characteristics of columella root cap can be performed easily under confocal microscopes. This simple technology, when used together with Lugol’s staining, provides a novel quantitative method to study the dynamics of stem cell behavior that govern homeostasis in the Arabidopsis columella root cap.
Highlights
In multicellular organisms, a pool of stem cells is maintained throughout the life cycle to produce exactly and only as many cells as necessary for a specific tissue, an astonishing natural phenomenon known as stem cell homeostasis
To permit quantitative analysis of stem cell homeostasis in the columella root cap, in this article, we propose a simple method to examine the rate of stem cell division and the progression of stem cell progeny differentiation in the Arabidopsis columella root cap
EdU Staining Reveals Disrupted Stem Cell Homeostasis in Mutants with Defects in the Columella Root Cap Using Lugol’s staining technology, accumulation of starch granules at the position of columella root cap stem cells was found after quiescent center (QC) ablation and in wox5-1 mutants (Figure 1B), resulting in the conclusion that the QC and WOX5 inhibit differentiation of columella root cap stem cells
Summary
A pool of stem cells is maintained throughout the life cycle to produce exactly and only as many cells as necessary for a specific tissue, an astonishing natural phenomenon known as stem cell homeostasis. Stem cells divide asymmetrically to produce two daughter cells of different fates. One of the daughters retains the self-renewal properties of the mother stem cell and keeps on dividing, whereas the other daughter is programmed to differentiate into a specialized mature cell. Asymmetrical division of stem cells and differentiation of mature cells are dynamically regulated so as to maintain the balance between self-renewal and lineage commitment in response to tissue requirements. To reveal the mechanisms that regulate stem cell homeostasis, the dynamics of stem cell behaviors have been studied in various animal tissues where rapid cell renewal is needed.
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