Knockout of the PKN Family of Rho Effector Kinases Reveals a Non-redundant Role for PKN2 in Developmental Mesoderm Expansion

Cell Rep. 2016 Jan 26;14(3):440-448. doi: 10.1016/j.celrep.2015.12.049. Epub 2016 Jan 7.

Abstract

In animals, the protein kinase C (PKC) family has expanded into diversely regulated subgroups, including the Rho family-responsive PKN kinases. Here, we describe knockouts of all three mouse PKN isoforms and reveal that PKN2 loss results in lethality at embryonic day 10 (E10), with associated cardiovascular and morphogenetic defects. The cardiovascular phenotype was not recapitulated by conditional deletion of PKN2 in endothelial cells or the developing heart. In contrast, inducible systemic deletion of PKN2 after E7 provoked collapse of the embryonic mesoderm. Furthermore, mouse embryonic fibroblasts, which arise from the embryonic mesoderm, depend on PKN2 for proliferation and motility. These cellular defects are reflected in vivo as dependence on PKN2 for mesoderm proliferation and neural crest migration. We conclude that failure of the mesoderm to expand in the absence of PKN2 compromises cardiovascular integrity and development, resulting in lethality.

Keywords: 4-OHT; 4-hydroxytamoxifen; ES cells; Embryonic stem cells; MEF; NCCs; PKC; PKN; Protein kinase N; mouse embryonic fibroblasts; neural crest cells; protein kinase C.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antineoplastic Agents / pharmacology
  • Apoptosis / drug effects
  • Cell Line
  • Cell Movement / drug effects
  • Cell Proliferation / drug effects
  • Embryo, Mammalian / cytology
  • Embryo, Mammalian / metabolism
  • Embryonic Development / drug effects
  • Genes, Reporter
  • Heart / growth & development
  • Mesoderm / cytology
  • Mesoderm / metabolism*
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Microscopy, Electron, Scanning
  • Myocardium / metabolism
  • Myocardium / pathology
  • Protein Kinase C / deficiency
  • Protein Kinase C / genetics*
  • Protein Kinase C / metabolism
  • Tamoxifen / analogs & derivatives
  • Tamoxifen / pharmacology

Substances

  • Antineoplastic Agents
  • Tamoxifen
  • afimoxifene
  • protein kinase N
  • Protein Kinase C