Establishment and characterization of a noradrenergic adrenal chromaffin cell line, tsAM5NE, immortalized with the temperature-sensitive SV40 T-antigen

Cell Biol Int. 2011 Apr;35(4):325-34. doi: 10.1042/CBI20090344.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adrenal Glands / cytology*
  • Animals
  • Antigens, Polyomavirus Transforming / genetics*
  • Cell Line, Tumor / cytology*
  • Cell Line, Tumor / metabolism
  • Cell Proliferation
  • Cells, Cultured
  • Chromaffin Cells / cytology*
  • Chromaffin Cells / metabolism
  • Glial Cell Line-Derived Neurotrophic Factor / metabolism
  • Mice
  • Mice, Transgenic
  • Nerve Growth Factors / metabolism
  • Neurogenesis
  • Neurons / cytology

Substances

  • Antigens, Polyomavirus Transforming
  • Glial Cell Line-Derived Neurotrophic Factor
  • Nerve Growth Factors