Intracellular localization of amyloid-β peptide in SH-SY5Y neuroblastoma cells.
Amyloid-β peptide (Aβ), the main component of Alzheimer's disease (AD) senile plaques, has been found to accumulate within the lysosomal compartment of AD neurons. We have previously shown that in differentiated SH-SY5Y neuroblastoma cells cultured under normal conditions, the majority of Aβ is localized extralysosomally, while oxidative stress significantly increases intralysosomal Aβ content through activation of macroautophagy. It is, however, not clear which cellular compartments contain extralysosomal Aβ in intact SH-SY5Y cells, and how oxidative stress influences the distribution of extralysosomal Aβ. Using confocal laser scanning microscopy and immunoelectron microscopy, we showed that in differentiated neuroblastoma cells cultured under normal conditions Aβ (Aβ40, Aβ42, and Aβ oligomers) is colocalized with both membrane-bound organelles (endoplasmic reticulum, Golgi complexes, multivesicular bodies/late endosomes, lysosomes, exocytotic vesicles and mitochondria) and non-membrane-bound cytosolic structures. Neuroblastoma cells stably transfected with AβPP Swedish KM670/671NL double mutation showed enlarged amount of Aβ colocalized with membrane compartments. Suppression of exocytosis by 5 nM tetanus toxin resulted in a significant increase of the amount of cytosolic Aβ as well as Aβ colocalized with exocytotic vesicles, endoplasmic reticulum, Golgi complexes, and lysosomes. Hyperoxia increased Aβ localization in the endoplasmic reticulum, Golgi apparatus, mitochondria, and lysosomes, but not in the secretory vesicles. These results indicate that in SH-SY5Y neuroblastoma cells intracellular Aβ is not preferentially localized to any particular organelle and, to a large extent, is secreted from the cells. Challenging cells to hyperoxia, exocytosis inhibition, or Aβ overproduction increased intracellular Aβ levels but did not dramatically changed its localization pattern.
KI-Alzheimer Disease Research Center, NVS, Novum, Karolinska Institutet, Stockholm, Sweden.
Lin
L
Zheng
Javier
J
Calvo-Garrido
Martin
M
Hallbeck
Kjell
K
Hultenby
Jan
J
Marcusson
Angel
A
Cedazo-Minguez
Alexei
A
Terman
D016229:Amyloid beta-Peptides / Q000032:analysis* / Q000378:metabolism*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000502:physiology; D006801:Humans; D007424:Intracellular Fluid / Q000737:chemistry* / Q000378:metabolism*; D009447:Neuroblastoma / Q000378:metabolism*
D016229:Amyloid beta-Peptides
Alzheimer's disease; amyloid β-protein; endosomes; exocytosis; lysosomes