Abstract
In CAA, the deposition of amyloid-β protein (Aβ) in blood vessels causes the death of cerebrovascular smooth muscle cells (CVSMCs), impairing the regulation of cerebral blood flow and increasing the risk of haemorrhage. It has been suggested that the death of CVSMCs depends on their uptake of Aβ. We describe advantages and limitations of a quantitative image-based method for measuring the uptake of fluorescently tagged Aβ by CVSMCs. This involves exposing CVSMCs for different periods to the tagged Aβ, fixing and mounting the cells and the use of image analysis software to quantify intracellular fluorescence. This method could of course be adapted for measuring the uptake of other proteins of interest. Although quantitative image-based analysis is more time-consuming than other, more widely used methods for measuring protein uptake, we show that it has important advantages in terms of allowing the exclusion of protein precipitates that adhere to but are not taken up by the CVSMCs. We suggest that this approach provides valuable additional information to flow cytometry and other automated methods for measurement of the uptake of Aβ into cells, and that researchers should include image-based analytical methods to avoid misinterpretation of data on protein uptake.
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MedCalc statistical software. www.medcalc.org
Acknowledgements
WAWR was supported by a grant from the Ministry of Higher Education and University of Malaya, Malaysia. This research was also supported by grants from Alzheimer’s Research UK and BRACE (Bristol Research into Alzheimer’s and Care of the Elderly).
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Ruzali, W.A.W., Love, S. (2015). Measurement of Aβ Uptake by Cerebrovascular Smooth Muscle Cells. In: Slevin, M., McDowell, G. (eds) Handbook of Vascular Biology Techniques. Springer, Dordrecht. https://doi.org/10.1007/978-94-017-9716-0_8
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DOI: https://doi.org/10.1007/978-94-017-9716-0_8
Publisher Name: Springer, Dordrecht
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