Premium
Novel functional interactions between Trk kinase and p75 neurotrophin receptor in neuroblastoma cells
Author(s) -
Lachyankar Mahesh B.,
Condon Peter J.,
Daou MarieClaire,
De Asit K.,
Levine John B.,
Obermeier Axel,
Ross Alonzo H.
Publication year - 2002
Publication title -
journal of neuroscience research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.72
H-Index - 160
eISSN - 1097-4547
pISSN - 0360-4012
DOI - 10.1002/jnr.10480
Subject(s) - nerve growth factor , tropomyosin receptor kinase a , low affinity nerve growth factor receptor , neurotrophin , microbiology and biotechnology , trk receptor , biology , epidermal growth factor , protein kinase b , pi3k/akt/mtor pathway , cellular differentiation , signal transduction , receptor , chemistry , biochemistry , gene
To understand the functional interactions between the TrkA and p75 nerve growth factor (NGF) receptors, we stably transfected LAN5 neuroblastoma cells with an expression vector for ET‐R, a chimeric receptor with the extracellular domain of the epidermal growth factor receptor (EGFR), and the TrkA transmembrane and intracellular domains. EGF activated the ET‐R kinase and induced partial differentiation. NGF, which can bind to endogenous p75, did not induce differentiation but enhanced the EGF‐induced response, leading to differentiation of almost all cells. A mutated NGF, 3T‐NGF, that binds to TrkA but not to p75 did not synergize with EGF. Enhancement of EGF‐induced differentiation required at least nanomolar concentrations of NGF, consistent with the low‐affinity p75 binding site. EGF may induce a limited number of neuronal cells because it also enhanced apoptosis. Both NGF and a caspase inhibitor reduced apoptosis and, thereby, enhanced differentiation. NGF seems to enhance survival through the phosphatidylinositol‐3 kinase (PI3K) pathway. Consistent with this hypothesis, Akt, a downstream effector of the PI3K pathway, was hyperphosphorylated in the presence of EGF+NGF. These results demonstrate that TrkA kinase initiates differentiation, and p75 enhances differentiation by rescuing differentiating cells from apoptosis via the PI3K pathway. Even though both EGF and NGF are required for differentiation of LAN5/ET‐R cells, only NGF is required for survival of the differentiated cells. In the absence of NGF, the cells die by an apoptotic mechanism, involving caspase‐3. An anti‐p75 antibody blocked the survival effect of NGF. Brain‐derived neurotrophic factor also enhanced cell survival, indicating that in differentiated cells, NGF acts through the p75 receptor to prevent apoptosis. © 2002 Wiley‐Liss, Inc.