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Published on: September 1, 2017
Sphk1 deficiency induces apoptosis and developmental defects and premature death in zebrafish
Ling Huang1, Fang Han1, Ying Huang1
1Key Laboratory of Healthy Mariculture for the East China Sea, Ministry of Agriculture and Rural Affairs, Jimei University, Xiamen, China.
Abstract:
The sphk1 gene plays a crucial role in cell growth and signal transduction. However, the developmental functions of the sphk1 gene during early vertebrate zebrafish embryo remain not completely understood. In this study, we constructed zebrafish sphk1 mutants through CRISPR/Cas9 to investigate its role in zebrafish embryonic development. Knockout of the sphk1 gene was found to cause abnormal development in zebrafish embryos, such as darkening and atrophy of the head, trunk deformities, pericardial edema, retarded yolk sac development, reduced heart rate, and premature death. The acetylcholinesterase activity was significantly increased after the knockout of sphk1, and some of the neurodevelopmental genes and neurotransmission system-related genes were expressed abnormally. The deletion of sphk1 led to abnormal expression of immune genes, as well as a significant decrease in the number of hematopoietic stem cells and neutrophils. The mRNA levels of cardiac development-related genes were significantly decreased. In addition, cell apoptosis increases in the sphk1 mutants, and the proliferation of head cells decreases. Therefore, our study has shown that the sphk1 is a key gene for zebrafish embryonic survival and regulation of organ development. It deepened our understanding of its physiological function. Our study lays the foundation for investigating the mechanism of the sphk1 gene in early zebrafish embryonic development.
Insights
The sphhingokinsae 1 (sphk1) gene is vital for zebrafish embryonic development. Its absence causes severe deformities, impacting organ development, immune function, and leading to premature death.
Area of Science:
- Developmental Biology
- Genetics
- Molecular Biology
Background:
- The sphhingokinsae 1 (sphk1) gene is known for its roles in cell growth and signal transduction.
- Its specific functions during early vertebrate embryonic development, particularly in zebrafish, are not fully elucidated.
Purpose of the Study:
- To investigate the developmental functions of the sphhingokinsae 1 (sphk1) gene in early zebrafish embryos.
- To characterize the phenotypic consequences of sphk1 gene knockout in zebrafish.
Main Methods:
- CRISPR/Cas9 gene editing was employed to create zebrafish sphk1 mutants.
- Phenotypic analysis, acetylcholinesterase activity assays, gene expression profiling, and cell apoptosis/proliferation assays were performed.
Main Results:
- Sphk1 knockout resulted in significant embryonic abnormalities including head deformities, trunk malformations, pericardial edema, yolk sac defects, reduced heart rate, and lethality.
- Increased acetylcholinesterase activity, abnormal expression of neurodevelopmental and neurotransmission genes, immune dysregulation, decreased hematopoietic stem cells and neutrophils, and reduced cardiac development gene expression were observed.
- Sphk1 mutants exhibited increased cell apoptosis and decreased head cell proliferation.
Conclusions:
- The sphhingokinsae 1 (sphk1) gene is essential for zebrafish embryonic survival and the proper regulation of organ development.
- This study enhances understanding of sphk1's physiological roles and provides a foundation for further mechanistic investigations in early zebrafish embryogenesis.
