Real-time imaging of glucocorticoid receptor dynamics in living neurons and glial cells in comparison with non-neural cells

Mayumi Nishi
Nobuyuki Takenaka
Noriyuki Morita
Takao Ito
Hitoshi Ozawa
Mitsuhiro Kawata

To investigate the intracellular trafficking of glucocorticoid receptor (GR) in response to various conditions in a single living cell
 a green fluorescent protein (GFP) and rat GR chimera construct (GFP-GR) was prepared. We transiently transfected GFP-GR into primary cultured rat hippocampal neurons
 cortical glial cells
 and non-neural cells
 e.g. COS-1 cells and CV-1 cells
 and compared the dynamic changes in subcellular localization of GFP-GR in these cells. When GFP-GR was expressed in the cells
 GFP-GR efficiently transactivated the mouse mammary tumour virus promoter in response to dexamethasone (DEX). The cytoplasm-to-nuclear translocation of GFP-GR induced with 10-7 M DEX
 a specific agonist of GR
 at 37°C was completed within 30 min in all cell types used
 and the rate of nuclear translocation was dependent on the ligand dose. The translocation of GFP-GR into the nucleus from the cytoplasm was induced in a ligand-specific manner
 similar to that of the native GR. The disruption of microtubules by colchicine or nocodazole showed no significant effect on the DEX-induced GFP-GR translocation from the cytoplasmic region to the nuclear region. The cells were not deteriorated during time-lapse imaging analysis for 1 h at 37 °C. The present findings suggest that the subcellular localization of GFP-GR is dynamically changed in response to extracellular and intracellular conditions
 and that there are no conspicuous variations in the manner of trafficking of GR among different types of cells in vitro.

European Journal of Neuroscience

11

6

10.1046/j.1460-9568.1999.00616.x

https://doi.org/10.1046/j.1460-9568.1999.00616.x
https://www.ncbi.nlm.nih.gov/pubmed/10336661