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Figure 8 | Molecular Neurodegeneration

Figure 8

From: Downregulation of CREB expression in Alzheimer's brain and in Aβ-treated rat hippocampal neurons

Figure 8

Neuroprotective effects of CREB. (A) Rat primary hippocampal neurons cultured on cover slips were transfected with green fluorescent protein (GFP) alone or in combination with wild type CREB or its mutants (KCREB or MCREB). The transfected neurons were exposed to Aβ (5 μM) for 48 h. The apoptotic cells are identified by nuclear condensation with DAPI (Blue). (B) Apoptosis among GFP expressing cells was quantitated by counting neurons with condensed nuclei. At least 500 GFP expressing cells were examined. Neurons were protected by CREB expression and CREB mutants exacerbated Aβ toxicity. *p < 0.01 compared to GFP control. (C) Rat primary hippocampal neurons cultured on cover slips were transduced with recombinant adenoviruses (AdV) encoding β galactosidase (β gal; control) or wild-type CREB (M.O.I: 50) for 48 h. The neurons were fixed, permeabilized, and immunostained for MAP2a (neurites) with FITC (green) and for CREB with Cy3 (red). (D) In addition, cultured rat primary hippocampal neurons were transduced with adenoviral β gal or CREB for 48 h. The whole cell lysates were prepared and immunoblotted for CREB and β galactosidase. Significant upregulation of CREB expression was seen following adenoviral transduction. (E) Cultured rat primary hippocampal neurons were transduced with adenoviruses encoding β galactosidase (β gal), wild type CREB (WT-CREB) or dominant negative mutant forms of CREB (KCREB or MCREB). After 24 h, neurons were exposed to 5 μM of Aβ fibrils for 48 h and then the activity of caspase-9, a marker for the intrinsic pathway of apoptosis was assayed. Significant protection of neurons from Aβ-induced apoptosis was observed. *p < 0.01 vs β galactosidase control.

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