501. RET PLCγ phosphotyrosine binding domain regulates Ca2+ signaling and neocortical neuronal migration.
- Author
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Lundgren TK, Nakahata K, Fritz N, Rebellato P, Zhang S, and Uhlén P
- Subjects
- Animals, Apoptosis, Blotting, Western, 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, Neurons metabolism, Phospholipase C gamma antagonists & inhibitors, Phospholipase C gamma genetics, Phosphorylation, Proto-Oncogene Proteins c-ret genetics, RNA, Messenger genetics, RNA, Small Interfering genetics, Real-Time Polymerase Chain Reaction, Calcium Signaling physiology, Cell Movement, Neocortex metabolism, Neurons cytology, Phospholipase C gamma metabolism, Phosphotyrosine metabolism, Proto-Oncogene Proteins c-ret metabolism
- 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.
- Published
- 2012
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