Figure 1.

Characterization of glial mesencephalic cultures from WT and parkin null mice after 20 DIV. Glial cultures were maintained in DMEM–FCS medium. A, Photomicrographs of glial cells staining for type 2 astrocytes, with anti-GFAP⁺, microglial cells (CD11b⁺), and nuclei (bis-benzimide) in PK-KO midbrain glial cultures. Scale bar, 30 μm. B, Immunodetection and densitometric analysis of astroglial (GFAP) protein by Western blot. C, Percentage of microglial cells present in WT and PK-KO cultures. Photomicrographs (D) and percentage of proliferating cells (BrdU⁺) and total nuclei (E) stained with bis-benzimide of WT and PK-KO cells present in glial cultures incubated, with BrdU for 24 h, 3 d after reseeding. Scale bar, 20 μm. F, Immunodetection and densitometric analysis of the proapoptotic and antiapoptotic proteins, Bclx_L/S present in WT and PK-KO glial cultures. Values for immunocytochemistry studies are expressed as the mean ± SEM from six replicates of four independent cultures. Values for Western blotting experiment are the mean ± SEM from four replicates of two independent cultures. Statistical analysis was performed by one-way ANOVA, followed by Newman–Keuls multiple comparison test. ⁺p < 0.05; ⁺⁺p < 0.01, PK-KO versus WT cultures.