Fig 1: Matrix metalloproteinases (MMPs) in vessels. Platelets were isolated from wildtype mice (a,c,e) or transgenic Alzheimer mice (b,d,f), labeled with the red fluorescent dye PKH26, transcardially infused, organotypic brain slices prepared, incubated for 2 weeks and then counterstained with Lectin-649 (shown in yellow) or with the MMP-2 or MMP-9 antibodies (Alexa-488, shown in green). As a negative control the primary antibody was omitted (a,b) showing no positive green staining. Note that red platelets only penetrated vessels when isolated from transgenic Alzheimer mice (b,d,f). Note that MMP-2 and MMP-9 were expressed only when platelets penetrated the vessel. Scale bar in (a) = 5 µm (a–f). In order to show release of MMP-2 and MMP-9, platelets (f) or cortical vessels (h) were isolated from wildtype or transgenic mice, incubated for 60 min and the levels of MMP-2 and MMP-9 measured by ELISA. Values are given as mean ± SEM (n = 10). Statistical analysis was performed by students T-Test (*p < 0.05). Note significant more release of MMP-2 and MMP-9 from platelets (g) isolated from transgenic mice but not from vessels (h).
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