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Development of transgenic Xenopus laevis with a high C-src gene expression
1Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Prague.
Molecular Reproduction and Development
|July 21, 1998
Summary
Elevated c-src expression in Xenopus laevis embryos, driven by Rous sarcoma virus (RSV) long terminal repeat (LTR), caused developmental abnormalities. The dosage of c-src correlated with RSV LTR integration, impacting embryogenesis.
Area of Science:
- Developmental Biology
- Molecular Biology
- Genetics
Background:
- The proto-oncogene c-src plays a role in cellular functions.
- Understanding the impact of oncogene dysregulation during embryonic development is crucial.
Purpose of the Study:
- To investigate the effects of elevated c-src expression on Xenopus laevis embryonic development.
- To determine the correlation between Rous sarcoma virus (RSV) long terminal repeat (LTR) integration and c-src expression levels.
Main Methods:
- Generation of transgenic Xenopus laevis expressing elevated c-src via mating with a male carrying integrated RSV LTR.
- Immunohistochemical analysis to detect c-Src dosage in larval tissues.
- Southern and polymerase chain reaction (PCR) analyses to confirm RSV LTR integration.
- RNA probing to quantify c-src mRNA levels.
Main Results:
- Offspring with higher c-src dosage exhibited abnormal development, including edema, head deformities, and defects in the eye and axial systems.
- A correlation was observed between the dosage of c-Src and the integrated RSV LTR.
- Aberrant larvae showed significantly higher c-src mRNA levels compared to controls.
Conclusions:
- Integrated RSV regulatory sequences can overstimulate c-src proto-oncogene expression in Xenopus laevis.
- Exceeding a specific c-src expression threshold interferes with normal embryonic development.
- Dosage-dependent effects of c-src highlight its critical role in early embryogenesis.