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. 2018 Oct 5;8(19):e3039.
doi: 10.21769/BioProtoc.3039.

Detection of Cell Death in Planarians

Nídia De Sousa  1   2 , Teresa Adell  1   2
Affiliations

Detection of Cell Death in Planarians

Nídia De Sousa et al. Bio Protoc. .

Abstract

Planarians are freshwater flatworms, well known for their ability to regenerate a complete organism from any piece of their body. Furthermore, planarians are constantly growing and degrowing throughout their lives, maintaining a functional and proportioned body. These properties rely on the presence of a population of adult stem cells and on the tight control of their cell renewal, which is based on the balance between the proliferation of new cells and their differentiation, and the death of unnecessary cells. Due to the importance of these two processes in planarian biology, over the years, researchers have optimized molecular techniques to detect both cell proliferation and cell death in planarians. Here, we present the two main protocols currently used for cell death detection and quantification in the planarian field: Caspase-3 activity quantification and TUNEL assay.

Keywords: Caspase-3 activity; Cell death; Cell turnover; Planarians; TUNEL assay.

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Conflict of interest statement

Competing interestsThe authors declare no conflicts of interest or competing interests.

Figures

Figure 1.
Figure 1.. Planarian homeostasis.
A. Planarians are able to grow and degrow during their lives, maintaining their body proportions and functionality. Image provided by Gustavo Rodriguez-Esteban. B. After a stimulus, proliferation and/or cell death can change in planarians. After feeding, neoblast proliferation increases throughout planarian body, and cell death is reduced to minimum levels, resulting in the increase of animal’s size. Conversely, when planarians are in starvation, neoblast proliferation is maintained at basal levels and cell death increases, which not only results in a decrease in body size but allows the reorganization of the tissues. Image from Nídia de Sousa Ph.D. thesis (de Sousa, 2017).
Figure 2.
Figure 2.. General view of Caspase-3 assay protocol.
A. Cartoon exemplifying protein extraction. B. Step-by-Step of protein concentration determination. C. Example of how load plate wells.
Figure 3.
Figure 3.. Result of a TUNEL assay.
The image shows the tail of a planarian imaged by confocal microscopy. White dots correspond to nucleus of dying cells. Scale bar = 250 μm.
See this image and copyright information in PMC

References

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