RET PLCγ phosphotyrosine binding domain regulates Ca2+ signaling and neocortical neuronal migration

PLoS One. 2012;7(2):e31258. doi: 10.1371/journal.pone.0031258. Epub 2012 Feb 15.

Abstract

The receptor tyrosine kinase RET plays an essential role during embryogenesis in regulating cell proliferation, differentiation, and migration. Upon glial cell line-derived neurotrophic factor (GDNF) stimulation, RET can trigger multiple intracellular signaling pathways that in concert activate various downstream effectors. Here we report that the RET receptor induces calcium (Ca(2+)) signaling and regulates neocortical neuronal progenitor migration through the Phospholipase-C gamma (PLCγ) binding domain Tyr1015. This signaling cascade releases Ca(2+) from the endoplasmic reticulum through the inositol 1,4,5-trisphosphate receptor and stimulates phosphorylation of ERK1/2 and CaMKII. A point mutation at Tyr1015 on RET or small interfering RNA gene silencing of PLCγ block the GDNF-induced signaling cascade. Delivery of the RET mutation to neuronal progenitors in the embryonic ventricular zone using in utero electroporation reveal that Tyr1015 is necessary for GDNF-stimulated migration of neurons to the cortical plate. These findings demonstrate a novel RET mediated signaling pathway that elevates cytosolic Ca(2+) and modulates neuronal migration in the developing neocortex through the PLCγ binding domain Tyr1015.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis
  • Blotting, Western
  • Calcium Signaling / physiology*
  • Cell Movement*
  • Cell Proliferation
  • Cells, Cultured
  • Embryo, Mammalian / cytology
  • Embryo, Mammalian / metabolism
  • Glial Cell Line-Derived Neurotrophic Factor / genetics
  • Glial Cell Line-Derived Neurotrophic Factor / metabolism
  • Humans
  • Immunoenzyme Techniques
  • Inositol 1,4,5-Trisphosphate Receptors / genetics
  • Inositol 1,4,5-Trisphosphate Receptors / metabolism
  • Mice
  • Neocortex / embryology
  • Neocortex / metabolism*
  • Neurons / cytology*
  • Neurons / metabolism
  • Phospholipase C gamma / antagonists & inhibitors
  • Phospholipase C gamma / genetics
  • Phospholipase C gamma / metabolism*
  • Phosphorylation
  • Phosphotyrosine / metabolism*
  • Proto-Oncogene Proteins c-ret / genetics
  • Proto-Oncogene Proteins c-ret / metabolism*
  • RNA, Messenger / genetics
  • RNA, Small Interfering / genetics
  • Real-Time Polymerase Chain Reaction

Substances

  • Glial Cell Line-Derived Neurotrophic Factor
  • Inositol 1,4,5-Trisphosphate Receptors
  • RNA, Messenger
  • RNA, Small Interfering
  • Phosphotyrosine
  • Proto-Oncogene Proteins c-ret
  • RET protein, human
  • Phospholipase C gamma