Figure 1 Immunocytochemical staining of a) neuronal cultures with anti-MAP2 antibody and b) astrocyte cultures with anti-GFAP antibody indicated neurons (red fluorescence) and astrocytes (green fluorescence) purities of greater than 90%, respectively. MAP: microtubule-associated … Effects

of Chronic Lithium Treatment on bcl-2 mRNA and Protein Fludarabine levels Quantitative real-time PCR was performed to evaluate the effect of lithium treatment on bcl-2 mRNA levels. Gel electrophoresis of the PCR products showed the specific amplification of a single 174bp amplicon, as was Inhibitors,research,lifescience,medical expected for the target (figure 2a). Lithium increased bcl-2 mRNA levels in the rat primary astrocyte, neuron, and mixed neuron-astrocyte cultures by factors of 1.59±0.08, 1.46±0.17, and 1.48±0.17, respectively, in comparison to their respective vehicle-treated cultures Inhibitors,research,lifescience,medical (figure 2a). However, these changes were not statistically significant (P=0.33, 0.64, and 0.57, respectively). In addition, there were no significant differences in the fold increases in bcl-2 mRNA levels of the lithium-treated cultures as compared with the

vehicle-treated cultures between the three different cultures. Figure 2 Effects of acute and chronic lithium treatment on bcl-2 a) mRNA and b) protein levels in rat primary astrocyte, neuronal, and mixed Inhibitors,research,lifescience,medical astrocyte-neuronal cultures treated with lithium (1 mM) or vehicle (100 µl sterile distilled water, control) for … Chronic treatment with lithium increased bcl-2 protein levels significantly in the astrocyte (175%; P<0.05) (figure 2b), but not in the neuronal (53%) or mixed neuron-astrocyte Inhibitors,research,lifescience,medical (30%) cultures in comparison to their respective vehicle-treated cultures. There were no significant differences in the basal level of bcl-2 protein (in the absence of lithium) (F=0.49, df=2; P=0.62) or percent change from basal bcl-2 protein level

(F=1.66, df=2; P=0.22) between the neuronal, astrocyte, and mixed cultures. Moreover, acute lithium treatment Inhibitors,research,lifescience,medical (1 mM for 24 h) had no significant effects on bcl-2 protein levels in the mixed neuron-astrocyte, neuron, and astrocyte cultures in comparison to the respective vehicle-treated cultures (P=0.89, 0.50, and 0.89, respectively) (figure 2b). Discussion The objective of the present study was to investigate below the effects of chronic lithium treatment on bcl-2 levels in three different cell type preparations enriched from rat primary cortical cultures. The findings indicate that chronic, but not acute, lithium treatment increased bcl-2 protein levels in the rat primary astrocyte cultures and exerted a trend increase in the neuronal culture. In contrast, neither acute nor chronic lithium treatment affected bcl-2 levels in the mixed neuron-astrocyte culture. To our knowledge, this study is the first of its kind to report that 7 days of lithium treatment (1 mM) increases bcl-2 protein levels in rat primary cortical astrocyte cultures.