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Determining Genetic Expression Profiles in C. elegans Using Microarray and Real-time PCR
Published on: July 30, 2011
Eimeria tenella: analysis of differentially expressed genes in the monensin- and maduramicin-resistant lines using
Tong Chen1, Wei Zhang, Jianmin Wang
1Department of Preventive Veterinary Medicine, College of Veterinary Medicine, China Agricultural University, 2 Yuan Ming Yuan Xi Road, Hai-dian District, Beijing 100094, PR China.
Abstract:
Drug resistance in coccidial populations has been a major problem to the prophylactic chemotherapy. Nevertheless, the mechanisms of the resistance are still poorly understood. In this report, cDNA array was designed based on the cDNA library for analysis of gene expression profile of the drug-resistant lines and their sensitive parental lines of Eimeria tenella. Two thousand eight hundred and six ESTs (expressed sequence tags) were obtained from 9600 clones which were randomly derived from the cDNA library with the 3' end sequencing. A total of 1424 TUTs (tentative unique transcripts) were determined from the database of our ESTs by bioinformatics analysis, from which a cDNA array was developed. The comparison of monensin-resistant line (MonR) and maduramicin-resistant line (MadR) with their sensitive parental lines was undertaken independently. It was observed that the number of the up-regulated genes was 5.58-fold more than that of the down-regulated genes in MonR when compared with its parental line. The up-regulated genes were mainly involved in cytoskeletal rearrangements and energy metabolism. In MadR, the number of the down-regulated genes was 3.07-fold more than the up-regulated genes, which were mainly related to invasion and cytoskeletal genes. However, in MadR the level of the glycometabolism-related and potential transporter genes were reduced. Our data suggest that the mechanisms of monensin and maduramicin resistance of E. tenella might be a very complex process.
Insights
Drug resistance in Eimeria tenella is a significant challenge. This study used cDNA arrays to analyze gene expression in drug-resistant lines, revealing complex mechanisms involving cytoskeletal and metabolic gene changes.
Area of Science:
- Veterinary Parasitology
- Molecular Biology
- Genomics
Background:
- Coccidial drug resistance poses a major challenge to effective chemotherapy in poultry.
- The underlying molecular mechanisms of resistance in Eimeria tenella remain poorly understood.
Purpose of the Study:
- To investigate the gene expression profiles of drug-resistant Eimeria tenella lines compared to their sensitive parental lines.
- To elucidate the complex mechanisms contributing to monensin and maduramicin resistance.
Main Methods:
- Development of a cDNA array based on expressed sequence tags (ESTs) from Eimeria tenella.
- Analysis of gene expression profiles in monensin-resistant (MonR) and maduramicin-resistant (MadR) lines versus sensitive parental lines using the cDNA array.
- Bioinformatic analysis of 2,806 ESTs to identify 1,424 tentative unique transcripts (TUTs).
Main Results:
- In MonR lines, upregulated genes (5.58-fold increase) were primarily associated with cytoskeletal rearrangements and energy metabolism.
- In MadR lines, downregulated genes (3.07-fold increase) were mainly related to invasion and cytoskeletal functions.
- Maduramicin resistance also involved reduced levels of glycometabolism and potential transporter genes.
Conclusions:
- The mechanisms of monensin and maduramicin resistance in Eimeria tenella are complex and involve distinct gene expression alterations.
- Understanding these genetic changes is crucial for developing strategies to combat drug resistance in coccidiosis.
- This study provides a foundation for further research into the molecular basis of antiparasitic drug resistance.
