Anales RANF

Effect of the β -amyloid peptide on microglia activation: ATP release @Real Academia Nacional de Farmacia. Spain 193 Figure 2. Effect of LPS treatment in the ATP release by the microglia. (A) Representative photomicrographs of quiescent (upper panels) and LPS treated (lower panels) microglial cells showing VNUT (green) and Iba1 (red) immunodetection. Nuclei are counterstained with DAPI (blue). A merge image is shown. Scale bar = 15 µm. (B, C) Luminometric measure of the extracellular ATP in microglial cells treated with LPS for 5 min (B) or 24 h (C). Luminiscence values were normalized with respect to those obtained in untreated (control) cells and are represented as a percentage. Data are the mean ± SEM of three independent experiments. **p < 0.01 (Student’s t -test). 3.3. Effect of the amyloid peptide β 1-42 in VNUT expression in microglial cells. Release of ATP Stimulation of the microglia with the amyloid peptide β 1-42 at a 5 µM concentration for 18h resulted in a significant increase in the ATP release to the extracellular medium when compared with the effect of the non- amyloidogenic (control) β 42-1 peptide (Figure 3A). However, VNUT expression was unaffected by the treatment with the amyloidogenic peptide as both the mRNA and protein levels of the transporter remained constant in β 1-42 or β 42-1 treated cells (Figura 3B). Moreover, the treatment with the amyloidogenic peptide induced a morphological change in the microglial cells which acquired the amoeboid morphology that is characteristic of the reactive microglia (Figure 3C). 0 50 100 150 200 ** Nomalized luminiscence vs Control (%) Control LPS 5min 0 50 100 150 Normalized luminiscence vs Control (%) Control LPS 24h B C VNUT DAPI VNUT Iba1 DAPI Iba1 DAPI CONTROL LPS A