CAS BI 525 Lecture Notes - Lecture 34: Amyloid, Gamma Secretase, Chinese Hamster Ovary Cell
17 Dec 2016
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Review: abeta oligomers, collected from supernatant of plate, left side: neurons. Tg: swedish mutation: right side: cultured mammalian cells. 7pa2 cells: cho cells transfected with swedish mutation (app) and gamma secretase cleavage site mutation: grow cells and collect supernatant (conditioned medium) then put it in a tube. Ip using antibody to target abeta (epitope) to separate it from the rest of conditioned medium. Next experiment (slide 18: created intracellularly and are secreted, microsomal preparation (intracellularly, show: oligomeric species are chemically soluble by treating with denaturing agents (urea and fa) the load does not change. 7pa2) rescue ltp: western blot ( e) shows that ide degrades monomers, not oligomers. Ide used to degrade monomers (not oligomers) from 7pa2 cells then administered to hippocampi slices, does not rescue ltp. Oligomers are the toxic forms to ltp. App is the same between 3 and 24 mo even though plaques increase.
