Related Experiment Video
Updated: Mar 8, 2026
Preparation of Mouse Embryonic Fibroblast Cells Suitable for Culturing Human Embryonic and Induced Pluripotent Stem Cells
Published on: June 21, 2012
PKN2 is essential for mouse embryonic development and proliferation of mouse fibroblasts
Sally Danno1, Koji Kubouchi1, Mona Mehruba1
1Graduate School of Medicine, Kobe University, Kobe, 650-0017, Japan.
Abstract:
PKN2, a member of the protein kinase N (PKN) family, has been suggested by in vitro culture cell experiments to bind to Rho/Rac GTPases and contributes to cell-cell contact and cell migration. To unravel the in vivo physiological function of PKN2, we targeted the PKN2 gene. Constitutive disruption of the mouse PKN2 gene resulted in growth retardation and lethality before embryonic day (E) 10.5. PKN2-/- embryo did not undergo axial turning and showed insufficient closure of the neural tube. Mouse embryonic fibroblasts (MEFs) derived from PKN2-/- embryos at E9.5 failed to grow. Cre-mediated ablation of PKN2 in PKN2flox/flox MEFs obtained from E14.5 embryos showed impaired cell proliferation, and cell cycle analysis of these MEFs showed a decrease in S-phase population. Our results show that PKN2 is essential for mouse embryonic development and cell-autonomous proliferation of primary MEFs in culture. Comparison of the PKN2-/- phenotype with the phenotypes of PKN1 and PKN3 knockout strains suggests that PKN2 has distinct nonredundant functions in vivo, despite the structural similarity and evolutionary relationship among the three isoforms.
Insights
Protein kinase N2 (PKN2) is crucial for embryonic development, impacting growth, neural tube closure, and cell proliferation. PKN2 knockout mice exhibit severe developmental defects and lethality, highlighting its essential in vivo functions.
Area of Science:
- Molecular Biology
- Developmental Biology
- Cell Biology
Background:
- Protein kinase N2 (PKN2) is implicated in cell-cell contact and migration via Rho/Rac GTPase interactions in vitro.
- The in vivo physiological functions of PKN2 remain largely uncharacterized.
Purpose of the Study:
- To investigate the in vivo physiological role of PKN2 during mouse embryonic development.
- To determine the necessity of PKN2 for cell-autonomous proliferation of mouse embryonic fibroblasts (MEFs).
Main Methods:
- Constitutive disruption of the mouse PKN2 gene.
- Analysis of PKN2 knockout (PKN2-/-) embryos for developmental abnormalities.
- Cre-mediated ablation of PKN2 in PKN2flox/flox MEFs.
- Cell proliferation assays and cell cycle analysis of MEFs.
Main Results:
- PKN2-/- embryos exhibited growth retardation and lethality before embryonic day 10.5.
- Developmental defects included failure of axial turning and insufficient neural tube closure.
- PKN2 deficiency in MEFs impaired cell proliferation, with a notable decrease in the S-phase population.
Conclusions:
- PKN2 is essential for mouse embryonic development, including axial turning and neural tube closure.
- PKN2 is required for cell-autonomous proliferation of primary MEFs.
- PKN2 possesses distinct, nonredundant in vivo functions compared to PKN1 and PKN3.
