We add amyloid β 1-42 to cells in order to create a model of Alzheimer's in cell culture. However, how can we determine exactly when Alzheimer's occurs in cells? Is there a method for this?
Hello Beyza Nur Ozkan ,
There are different ways to assess whether your amyloid peptides are actually affecting your cells.
1) Cell death (most used method with administration of exogenous amyloid beta peptides). Amyloid beta 1-42 is know to trigger cell death in culture so commonly used protocols and kits to check cell death, apoptosis or proliferation could be useful in this case. These are often colorimetric or fluorimetric assays so really easy to perform.
2) Mitochondrial homeostasis. Amyloid beta 1-42 triggers many defects in mitochondria. First, you could check mitochondrial morphology (mitochondria should appear less interconnected and more rounded after exposure to amyloid beta peptide). Second, you could check mitochondrial function, ATP production and oxygen consumption should be decreased for example.
3) Gene expression. According to the cell model you are investigating you could check expression of different pathways whose upregulation would indicate insurgence of proteotoxic stress: heat shock response, inflammation, apoptosis, proteosome, unfolded protein response, etc.
Hope this help you.
Good luck,
Mario
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