Subcellular localization of glucocorticoid receptor protein in the human kidney glomerulus.
BACKGROUND
The detailed mechanisms of glucocorticoid action in idiopathic nephrotic syndrome and progressive glomerulonephritides have not been clearly elucidated. The pharmacological actions of glucocorticoids are mediated by their binding to an intracellular protein, the glucocorticoid receptor (GR). The determination of GR localization in normal glomerular cells is essential to elucidate the mechanisms of glucocorticoid action in various glomerular diseases.


METHODS
We carried out an immunoblot examination using antihuman GR-specific antibody and homogenates of isolated normal human glomeruli and mesangial cells in culture. Immunohistochemical examinations were also performed on normal human kidney specimens at light and electron microscopic levels. The nuclear translocation of GRs elicited by ligand binding was further investigated by confocal laser-scanning microscopic inspection of freshly isolated glomeruli and mesangial cells cultured with dexamethasone.


RESULTS
An immunoblot examination demonstrated the presence of a 94 kDa protein, a molecular weight consistent with that of GRs, in the homogenates of glomeruli and cultured mesangial cells. By light microscopic examination, GRs were strongly detected in the nucleus and moderately in the cytoplasm of all glomerular cells, parietal and visceral epithelial cells, endothelial cells, and mesangial cells. By electron microscopic examination, the nuclear GRs of all glomerular cells were found to be diffusely distributed in the euchromatin. Additionally, the immunofluorescence intensities of nuclear GRs in isolated glomeruli and mesangial cells in culture became more intense by the addition of dexamethasone.


CONCLUSIONS
Our findings suggest that all subsets of human glomerular cells definitely express the GR protein, which potentially undergoes translocation by glucocorticoids.
Department of Pediatrics, Kyorin University School of Medicine, Tokyo, Japan. yang@kt.rim.or.jp
K
K
Yan
A
A
Kudo
H
H
Hirano
T
T
Watanabe
T
T
Tasaka
S
S
Kataoka
N
N
Nakajima
Y
Y
Nishibori
T
T
Shibata
T
T
Kohsaka
E
E
Higashihara
H
H
Tanaka
H
H
Watanabe
T
T
Nagasawa
S
S
Awa
D002467:Cell Nucleus / Q000378:metabolism / Q000648:ultrastructure; D002478:Cells, Cultured; D003907:Dexamethasone / Q000494:pharmacology; D019084:Fluorescent Antibody Technique, Indirect; D005920:Glomerular Mesangium / Q000166:cytology / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D005938:Glucocorticoids / Q000494:pharmacology; D006801:Humans; D015151:Immunoblotting; D007678:Kidney Glomerulus / Q000166:cytology / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D016253:Microscopy, Immunoelectron; D011965:Receptors, Glucocorticoid / Q000378:metabolism*; D014018:Tissue Distribution / Q000502:physiology
D005938:Glucocorticoids; D011965:Receptors, Glucocorticoid; D003907:Dexamethasone