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Updated: Jun 13, 2026
Stem cell-like Xenopus Embryonic Explants to Study Early Neural Developmental Features In Vitro and In Vivo
Published on: February 2, 2016
Identification, characterization, and effects of Xenopus laevis PNAS-4 gene on embryonic development
Fei Yan1, Xu-zhi Ruan, Han-shuo Yang
1State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, West China Medical School, Sichuan University, Chengdu 610041, China.
Abstract:
Apoptosis plays an important role in embryonic development. PNAS-4 has been demonstrated to induce apoptosis in several cancer cells. In this study, we cloned Xenopus laevis PNAS-4 (xPNAS-4), which is homologous to the human PNAS-4 gene. Bioinformatics analysis for PNAS-4 indicated that xPNAS-4 shared 87.6% identity with human PNAS-4 and 85.5% with mouse PNAS-4. The phylogenetic tree of PNAS-4 protein was also summarized. An analysis of cellular localization using an EGFP-fused protein demonstrated that xPNAS-4 was localized in the perinuclear region of the cytoplasm. RT-PCR analysis revealed that xPNAS-4, as a maternally expressed gene, was present in all stages of early embryo development. Whole-mount in situ hybridization showed that xPNAS-4 was mainly expressed in ectoderm and mesoderm. Furthermore, microinjection of xPNAS-4 mRNA in vivo caused developmental defects manifesting as a small eye phenotype in the Xenopous embryos, and as a small eye or one-eye phenotype in developing zebrafish embryos. In addition, embryos microinjected with xPNAS-4 antisense morpholino oligonucleotides (MOs) exhibited a failure of head development and shortened axis.
Insights
Xenopus laevis PNAS-4 (xPNAS-4) is a maternally expressed gene crucial for embryonic development. Its microinjection causes developmental defects, including small eyes and head axis shortening, highlighting its role in embryogenesis.
Area of Science:
- Developmental Biology
- Molecular Biology
- Genetics
Background:
- Apoptosis is vital for embryonic development.
- PNAS-4 induces apoptosis in cancer cells.
- xPNAS-4 is homologous to human PNAS-4.
Purpose of the Study:
- To clone and characterize Xenopus laevis PNAS-4 (xPNAS-4).
- To investigate the role of xPNAS-4 in early embryonic development.
- To analyze the effects of xPNAS-4 manipulation on embryonic development.
Main Methods:
- Cloning of xPNAS-4.
- Bioinformatics analysis for sequence identity and phylogenetic tree construction.
- Cellular localization using EGFP-fusion protein.
- RT-PCR and whole-mount in situ hybridization for expression analysis.
- Microinjection of xPNAS-4 mRNA and antisense morpholino oligonucleotides (MOs).
Main Results:
- xPNAS-4 shares high sequence identity with human and mouse PNAS-4.
- xPNAS-4 localizes to the perinuclear region of the cytoplasm.
- xPNAS-4 is a maternally expressed gene present throughout early embryonic development, primarily in ectoderm and mesoderm.
- Microinjection of xPNAS-4 mRNA causes small eye phenotypes in Xenopus and zebrafish embryos.
- xPNAS-4 MOs lead to head development failure and shortened axes.
Conclusions:
- xPNAS-4 plays a significant role in embryonic development.
- Dysregulation of xPNAS-4 leads to severe developmental abnormalities.
- xPNAS-4 is a conserved gene with critical functions in vertebrate embryogenesis.
