期刊
BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES
卷 1860, 期 11, 页码 2204-2214出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.bbamem.2018.08.010
关键词
Amyloid-beta; Endocytosis; Alzheimer's disease; Protein aggregation; Cell surface proteoglycan; Chinese hamster ovary cells
资金
- Wenner-Gren Foundations
- Swedish Research Council [2016-03902]
- Swedish Innovation Agency [2011-03488]
- Linnaeus Centre SUPRA
- Magnus Bergvall Foundations
- IngaBritt foundation
- Arne Lundberg foundation
Proteoglycans (PGs) have been found in Alzheimer's disease amyloid-beta (A beta) plaques and their glycosaminoglycan chains reportedly influence A beta aggregation, neurotoxicity and intracellular accumulation in cell and animal models, but their exact pathophysiological role(s) remain unclear. We have studied the cellular uptake of fluorescently labelled A beta(1-42) and A beta(1-40) peptides in normal CHO cells (K1) and the mutant cell line (pgsA-745) which lacks all protein-attached heparan and chondroitin sulfate chains. After 24 h of incubation, CHO-K1 accumulates more A beta(1-42) and A beta(1-40) compared with CHO-pgsA-745, consistent with the suggested role of PGs in A beta uptake. However, after short incubation times (<= 3 h) there was no difference; moreover, the time evolution of A beta(1-42) accumulation in CHO-K1 followed an unusual sigmoidal-like trend, indicating a possible involvement of PG-mediated peptide aggregation in A beta endocytosis. Neither A beta(1-42) nor A beta(1-40) could stimulate uptake of a 10 kDa dextran (a general endocytosis marker) suggesting that A beta-induced upregulation of endocytosis does not occur. CHO-K1 cells contained a higher number of A beta(1-42)-positive vesicles, but the intensity difference per vesicle was only marginal suggesting that the superior accumulation of A beta(1-42) stems from a higher number of endocytic events. FRET imaging support that intracellular A beta(1-42) is aggregated in both cell types. We also report that CHO-pgsA-745 cells perform less endocytosis than CHO-K1 and, albeit this does not explain their difference in A beta internalisation, we discuss a general method for data compensation. Altogether, this study contributes new insights into the mechanisms of PG-mediated A beta uptake that may be relevant for our understanding of their role in AD pathology.
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