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Updated: May 29, 2026
Culturing and Genetically Manipulating Entomopathogenic Nematodes
Published on: March 31, 2022
Caspase dependent programmed cell death in developing embryos: a potential target for therapeutic intervention
Alok Das Mohapatra1, Sunil Kumar, Ashok Kumar Satapathy
1Department of Infectious Disease Biology, Institute of Life Sciences, DBT, Ministry of Science and Technology, Government of India, Bhubaneswar, India.
Background:
Successful embryogenesis is a critical rate limiting step for the survival and transmission of parasitic worms as well as pathology mediated by them. Hence, blockage of this important process through therapeutic induction of apoptosis in their embryonic stages offers promise for developing effective anti-parasitic measures against these extra cellular parasites. However, unlike in the case of protozoan parasites, induction of apoptosis as a therapeutic approach is yet to be explored against metazoan helminth parasites.
Methodology/Principal Findings:
For the first time, here we developed and evaluated flow cytometry based assays to assess several conserved features of apoptosis in developing embryos of a pathogenic filarial nematode Setaria digitata, in-vitro as well as ex-vivo. We validated programmed cell death in developing embryos by using immuno-fluorescence microscopy and scoring expression profile of nematode specific proteins related to apoptosis [e.g. CED-3, CED-4 and CED-9]. Mechanistically, apoptotic death of embryonic stages was found to be a caspase dependent phenomenon mediated primarily through induction of intracellular ROS. The apoptogenicity of some pharmacological compounds viz. DEC, Chloroquine, Primaquine and Curcumin were also evaluated. Curcumin was found to be the most effective pharmacological agent followed by Primaquine while Chloroquine displayed minimal effect and DEC had no demonstrable effect. Further, demonstration of induction of apoptosis in embryonic stages by lipid peroxidation products [molecules commonly associated with inflammatory responses in filarial disease] and demonstration of in-situ apoptosis of developing embryos in adult parasites in a natural bovine model of filariasis have offered a framework to understand anti-fecundity host immunity operational against parasitic helminths.
Conclusions/Significance:
Our observations have revealed for the first time, that induction of apoptosis in developing embryos can be a potential approach for therapeutic intervention against pathogenic nematodes and flow cytometry can be used to address different issues of biological importance during embryogenesis of parasitic worms.
Insights
Inducing apoptosis in parasitic worm embryos offers a new therapeutic strategy. Flow cytometry assays effectively measure programmed cell death in nematode embryos, with curcumin showing promise as an anti-parasitic agent.
Area of Science:
- Parasitology
- Developmental Biology
- Cell Biology
Background:
- Embryogenesis is crucial for parasitic worm survival and pathology.
- Targeting embryogenesis via apoptosis offers a novel anti-parasitic strategy.
- Apoptosis induction for helminth parasites remains largely unexplored.
Purpose of the Study:
- To develop and validate flow cytometry assays for apoptosis in parasitic nematode embryos.
- To investigate the mechanisms of apoptosis in Setaria digitata embryos.
- To evaluate the efficacy of pharmacological compounds in inducing apoptosis.
Main Methods:
- Developed flow cytometry assays for apoptosis in Setaria digitata embryos.
- Validated programmed cell death using immunofluorescence and apoptosis-related protein expression.
- Assessed caspase dependency and reactive oxygen species (ROS) involvement.
- Evaluated pharmacological agents including curcumin, primaquine, and chloroquine.
Main Results:
- Established flow cytometry assays for assessing apoptosis in nematode embryos.
- Confirmed apoptosis as a caspase-dependent process mediated by ROS.
- Curcumin demonstrated the highest efficacy in inducing apoptosis, followed by primaquine.
- Demonstrated in-situ apoptosis in adult parasites from a bovine model.
Conclusions:
- Apoptosis induction in developing embryos is a viable therapeutic approach against nematodes.
- Flow cytometry is a valuable tool for studying embryogenesis in parasitic worms.
- Findings provide a framework for understanding anti-fecundity immunity in helminth infections.
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