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Establishment and characterization of a noradrenergic adrenal chromaffin cell line, tsAM5NE, immortalized with the temperature‐sensitive SV40 T‐antigen
Author(s) -
Kohno Susumu,
Murata Tomiyasu,
Koide Naoshi,
Hikita Kiyomi,
Kaneda Norio
Publication year - 2011
Publication title -
cell biology international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.932
H-Index - 77
eISSN - 1095-8355
pISSN - 1065-6995
DOI - 10.1042/cbi20090344
Subject(s) - glial cell line derived neurotrophic factor , neurturin , gdnf family of ligands , neurotrophic factors , chromaffin cell , cell culture , biology , microbiology and biotechnology , cellular differentiation , adrenal medulla , endocrinology , catecholamine , receptor , gene , biochemistry , genetics
We established a clonal adrenal medullary cell line, named tsAM5NE, from transgenic mice harbouring the temperature‐sensitive Simian virus 40 large T‐antigen gene, under the control of the tyrosine hydroxylase promoter. tsAM5NE cells conditionally grew at a permissive temperature of 33°C and exhibited the noradrenergic chromaffin cell phenotype. To understand the characteristics of tsAM5NE cells, we first examined the responsiveness of the cells to ligands of the GDNF (glial cell line‐derived neurotrophic factor) family. tsAM5NE cells proliferated at the permissive temperature of 33°C in response to either GDNF or neurturin, but not artemin or persephin. At the non‐permissive temperature of 39°C, GDNF or neurturin caused tsAM5NE cells to differentiate into neuron‐like cells; however, the differentiated cells died in a time‐dependent manner. Interestingly, LIF (leukaemia inhibitory factor) 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 LIF, the morphological change induced by the temperature shift was associated with up‐regulated expression of neuronal markers, indicating that the cells had indeed undergone neuronal differentiation. Thus, we demonstrated that tsAM5NE cells had the capacity to terminally differentiate into neuron‐like cells in response to GDNF plus LIF when the oncogene was inactivated by the temperature shift. Thus, this cell line provides a useful model system for studying the mechanisms regulating neuronal differentiation.