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Neuronal differentiation elicited by glial cell line‐derived neurotrophic factor and ciliary neurotrophic factor in adrenal chromaffin cell line tsAM5D immortalized with temperature‐sensitive SV40 T‐antigen
Author(s) -
Murata Tomiyasu,
Tsuboi Masaru,
Koide Naoshi,
Hikita Kiyomi,
Kohno Susumu,
Kaneda Norio
Publication year - 2008
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.21632
Subject(s) - ciliary neurotrophic factor , neurotrophic factors , chromaffin cell , glial cell line derived neurotrophic factor , cell culture , gdnf family of ligands , neuroscience , microbiology and biotechnology , nerve growth factor , neurotrophin , line (geometry) , biology , immunology , medicine , adrenal medulla , catecholamine , genetics , receptor , geometry , mathematics
To understand the characteristics of tsAM5D cells immortalized with the temperature‐sensitive simian virus 40 large T‐antigen, we first examined the responsiveness of the cells to ligands of the glial cell line‐derived neurotrophic factor (GDNF) family. tsAM5D cells proliferated at the permissive temperature of 33°C in response to either GDNF or neurturin, but not persephin or artemin. At the nonpermissive temperature of 39°C, GDNF or neurturin caused tsAM5D cells to differentiate into neuron‐like cells; however, the differentiated cells died in a time‐dependent manner. Interestingly, ciliary neurotrophic factor (CNTF) did not affect the GDNF‐mediated cell proliferation at 33°C but promoted the survival and differentiation of GDNF‐treated cells at 39°C. In the presence of GDNF plus CNTF, the morphological change induced by the temperature shift was associated with up‐regulated expression of various neuronal marker genes, indicating that the cells had undergone neuronal differentiation. In addition, tsAM5D cells caused to differentiate by GDNF plus CNTF at 39°C became dependent solely on nerve growth factor (NGF) for their survival and neurite outgrowth. Moreover, upon treatment with GDNF plus CNTF, the dopaminergic phenotype was suppressed by the temperature shift. Thus, we demonstrated that tsAM5D cells had the capacity to differentiate terminally into neuron‐like cells in response to GDNF plus CNTF when the oncogene was inactivated by the temperature shift. This cell line provides a useful model system for studying the role of a variety of signaling molecules for GDNF/CNTF‐induced neuronal differentiation. © 2008 Wiley‐Liss, Inc.