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Identification of apoptosis-related genes and transcription variations in response to microcystin-LR in zebrafish
Lili Wei1, Dave Hoole2, Baojian Sun3
1College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, Jiangxi Province, China State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, China.
Abstract:
There is growing evidence that the effects of microcystin-LR (MC-LR) are closely related to apoptosis. This study utilized microarray to identify the apoptosis-related genes induced by MC-LR in zebrafish liver. The messenger RNA abundance of some apoptosis-related genes was found to be increased, including five tumor necrosis factor (TNF)-related members (apoptosis regulatory protein siva, tumor necrosis factor-α (tnfa) TNF (ligand) superfamily member 10 (tnfsf10), TNF-inducible protein 6 (tnfaip6) and TNF receptor associated factor 2 binding protein (traf2bp)), three p53-related genes (tumor protein p53 inducible nuclear protein 1 (tp53inp1), p53-induced protein phosphatase 1 (ppm1d) and a novel apoptosis stimulating protein of p53 (aspp2)), bcl 2 family members (proapoptosis gene bax and antiapoptosis gene mcl 1), caspases (caspase y (caspy) and a PYD and CARD domain-containing protein (pycard)) and the transforming growth factor beta (TGF-β) induced apoptosis protein 2 (taip2). Real-time polymerase chain reaction was used to study the kinetic transcriptional changes in seven apoptosis-related genes. Elevated transcription of p53, tp53inp1, mcl 1 and taip2 could only be detected at 6 h, increased transcription of the antagonist molecules, bcl 2 and bax could be detected at most time points and the significant change of caspy could be found at 48 h and 72 h after stimulation. Taken together, the results obtained in the present study clearly demonstrate that large amount of apoptosis-related genes are involved in the regulation of MC-LR-induced apoptosis.
Insights
Microcystin-LR (MC-LR) exposure induces apoptosis by upregulating numerous apoptosis-related genes in zebrafish liver. Key genes involved include tumor necrosis factor (TNF) members, p53-related factors, and caspases.
Area of Science:
- Environmental toxicology
- Molecular biology
- Cellular apoptosis
Background:
- Microcystin-LR (MC-LR) is a potent hepatotoxin linked to apoptosis.
- Understanding the molecular mechanisms of MC-LR-induced apoptosis is crucial for risk assessment.
Purpose of the Study:
- To identify apoptosis-related genes affected by MC-LR exposure in zebrafish liver.
- To analyze the temporal expression patterns of these genes.
Main Methods:
- Microarray analysis to screen for differentially expressed genes.
- Real-time polymerase chain reaction (PCR) to validate gene expression kinetics.
Main Results:
- MC-LR significantly altered the expression of multiple apoptosis-related genes, including TNF-related members, p53-related genes, bcl-2 family members, caspases, and TGF-β-induced apoptosis protein 2 (taip2).
- Kinetic analysis revealed distinct temporal expression profiles for genes like p53, tp53inp1, mcl-1, taip2, bcl-2, bax, and caspase y (caspy).
Conclusions:
- MC-LR induces apoptosis through the complex regulation of a wide array of apoptosis-related genes in zebrafish.
- The findings provide insights into the molecular pathways mediating MC-LR toxicity.
