Phosphorylation status of glucocorticoid receptor, heat shock protein 70, cytochrome c and Bax in lymphocytes of euthymic, depressed and manic bipolar patients

Bipolar disorder (BD), a severe mental illness, has been correlated with alterations in glucocorticoid receptor (GR) signaling. Since it is phosphorylated GR that contributes to receptor function and determines its transcriptional activity, the Ser211 being a biomarker for activated GR in vivo, it is pertinent that we seek to determine the putative role of the total phosphorylation status of GR and site-specific phosphorylation at serine 211 (S211) in BD and their possible association with parameters of apoptosis. In lymphocytes from 48 BD patients under multiple psychotropic therapy and 20 healthy subjects, we measured whole cell GR, total GR phosphorylation, and phosphorylation of GR at serine 211 in nucleus, using immunoprecipitation, phosphospecific antibody and Western-blot analysis. Cytosolic cytochrome c and Bax and whole cell HSP70 were determined by immunoblot analysis. One-way ANOVA statistical analysis was carried out. Total phosphorylated GR was lower (P < 0.001) while the GR S211 was higher (P < 0.001) in all BD patients as compared to healthy subjects. HSP70 was reduced in euthymic (P < 0.05), depressed (P < 0.001) and manic (P < 0.001) as compared to healthy subjects. Cytochrome c was higher in all-patient groups as compared to healthy subjects, however without reaching statistical significance (P > 0.05). Bax levels were lower in the cytosolic fraction of all three BD groups. We provide the first evidence of altered GR phosphorylation joined with signs of apoptosis in lymphocytes of BD patients and suggest that the phosphorylation status of GR may play a role in the pathophysiology of bipolar disorder. © 2009 Elsevier Ltd. All rights reserved

Peripheral blood lymphocytes from patients with bipolar disorder demonstrate apoptosis and differential regulation of advanced glycation end products and s100b

Background: This study addresses the expression of the glycosylated proteins known as advanced glycation end products (AGEs), the calcium binding protein S100B and the apoptotic parameters cytochome c and caspase-3 activity in peripheral lymphocyte cytosolic extracts from a sample of bipolar disorder (BD) patients and healthy (control) subjects. Methods: Cross-sectional study of 35 patients with a clinical diagnosis of bipolar disease (10 euthymic, 12 depressed, 13 manic) and 10 healthy control subjects. Lymphocytes were used as a surrogate model in BD diagnosis and treatment. AGEs and S100B in lymphocyte cell extracts were measured by commercially available enzyme-linked immunosorbent assay. Results: AGEs were lower in all BD patients compared to healthy subjects. Depressed patients had approximately two-fold higher S100B levels compared to healthy subjects. Manic and depressed BD patients had increased superoxide dismutase mRNA levels. Apoptosis as measured by BAX/Bcl2 ratio, cytochrome c release, caspase-3 activity was increased in manic and depressed patients compared to healthy subjects. In the depressed patients, S100B levels correlated with cytochrome c release. Conclusions: In conclusion, our study shows decreased AGEs and increased S100B levels and caspase downstream apoptosis in peripheral lymphocytes of BD patients that may underlie disease etiopathogenesis