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A protocol describing pharynx counts and a review of other assays of apoptotic cell death in the nematode worm Caenorhabditis elegans

Nature Protocols volume 2, pages 705–714 (2007)Cite this article

Abstract

Studies of the nematode worm Caenorhabditis elegans have provided important insights into the genetics of programmed cell death (PCD), and revealed molecular mechanisms conserved from nematodes to humans. The organism continues to offer opportunities to investigate the processes of apoptosis under very well-defined conditions and at single-cell resolution in living animals. Here, a survey of the common methods used to study the process of PCD in C. elegans is described. Detailed instructions are provided for one standard method—the counting of extra cells of the anterior pharynx—a quantitative technique that can be used to detect even very subtle alterations in the progression of apoptotic cell death.

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Figure 1: A conserved core pathway for the execution of programmed cell death.
Figure 2: CED-1∷GFP surrounds apoptotic cell corpses.
Figure 3: Observation of persistent cell corpses using Nomarksi microscopy.
Figure 4: Using Nomarski microscopy to score PCD in the postdeirid.
Figure 5: Using Nomarski microscopy to identify cells sexually dimorphic for PCD.
Figure 6: Diagrams illustrating the preparation of agarose pads for mounting C. elegans for Nomarski microscopy.
Figure 7: Positions of nuclei in the wild-type anterior phanyx.
Figure 8: An example of nuclear positions in the anterior phanynx of a cell-death-defective animal.

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Acknowledgements

I thank H. Robert Horvitz, Brendan Galvin and Daniel Denning for their comments on the manuscript, Brian Harfe and Andrew Fire for providing the unpublished egl-17∷gfp reporter and information on its expression in the M4 neuron, Brendan Galvin for discussions regarding the use of lin-11∷gfp, Maureen Barr and Paul Sternberg for pkd-2∷gfp, Robyn Lints and Scott Emmons for cat-2∷gfp, Catherine Branda and Michael Stern for egl-17∷gfp and Mike Hurwitz for the ced-1(e1735); bcIs39 strain. This work was supported by a David H. Koch Graduate Fellowship and by funding from the Howard Hughes Medical Institute to H. Robert Horvitz.

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    Hillel T Schwartz

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Schwartz, H. A protocol describing pharynx counts and a review of other assays of apoptotic cell death in the nematode worm Caenorhabditis elegans. Nat Protoc 2, 705–714 (2007). https://doi.org/10.1038/nprot.2007.93

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