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Published on: August 19, 2014
Programmed cell death in procyclic form Trypanosoma brucei rhodesiense --identification of differentially expressed
S C Welburn1, S Lillico, N B Murphy
1Centre for Tropical Veterinary Medicine, University of Edinburgh, Scotland. sue.welburn@tsetse.demon.co.uk
Abstract:
Trypanosoma brucei rhodesiense can be induced to undergo apoptosis after stimulation with Con A. As cell death in these parasites is associated with de novo gene expression we have applied a differential display technique, Randomly Amplified Differential Expressed Sequence-Polymerase Chain Reaction (RADES-PCR) to the study of gene expression during Con A induced cell death in these organisms. Twenty-two differentially displayed products have been cloned and sequenced. These represent the first endogenous genes to be identified as implicated in cellular death in trypanosomatids (the most primitive eukaryote in which apoptosis has been described). Evidence for an ancestral death machinery, 'proto-apoptosis' in single celled organisms is discussed.
Insights
Researchers identified genes involved in programmed cell death in Trypanosoma brucei rhodesiense using Randomly Amplified Differential Expressed Sequence-Polymerase Chain Reaction (RADES-PCR). This study reveals key genetic factors in apoptosis for these primitive eukaryotes.
Area of Science:
- Molecular Biology
- Parasitology
- Eukaryotic Cell Biology
Background:
- Apoptosis, or programmed cell death, is a fundamental biological process observed across eukaryotes.
- Trypanosomes, including Trypanosoma brucei rhodesiense, are single-celled eukaryotes with a complex life cycle and susceptibility to induced cell death.
- Gene expression changes are known to accompany cell death in these parasites.
Purpose of the Study:
- To investigate gene expression changes during Concanavalin A (Con A)-induced apoptosis in Trypanosoma brucei rhodesiense.
- To identify novel endogenous genes implicated in the cellular death pathway of trypanosomatids.
- To explore the evolutionary origins of apoptosis in primitive eukaryotes.
Main Methods:
- Induction of apoptosis in Trypanosoma brucei rhodesiense using Concanavalin A (Con A).
- Application of Randomly Amplified Differential Expressed Sequence-Polymerase Chain Reaction (RADES-PCR) for differential gene display.
- Cloning and sequencing of differentially expressed cDNA products.
Main Results:
- Twenty-two differentially expressed sequence tags were identified during Con A-induced cell death.
- These represent the first endogenous genes linked to apoptosis in trypanosomatids.
- The findings suggest a conserved, ancestral 'proto-apoptosis' mechanism in single-celled organisms.
Conclusions:
- Gene expression plays a crucial role in Con A-induced apoptosis in Trypanosoma brucei rhodesiense.
- RADES-PCR is an effective method for identifying novel genes involved in parasite cell death.
- The study provides evidence for an ancient evolutionary basis of apoptotic pathways in eukaryotes.
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