Friday, March 13, 2009

Amyloid Beta and E18 Primary Hippocampal Neurons

We would like to feature an interesting application of our E18 Rat Primary Hippocampal Neurons.

Related Publication:

Karunya K. Kandimalla1, Olenych G. Scott, Smita Fulzele1, Michael W. Davidson, Joseph F. Poduslo. Mechanism of Neuronal versus Endothelial Cell Uptake of Alzheimer's Disease Amyloid β Protein. PLoS ONE 4(2): e4627. doi:10.1371/journal.pone.0004627.

... Rat primary hippocampal (RPH) neurons were isolated from the hippocampii of 18-day-old embryonic Sprague Dawley rat brains (Neuromics, Edina, MN). The hippocampii were dispersed using a fire polished Pasteur pipette and plated on poly-D-lysine (Sigma-Aldrich, St. Louis, MO) coated glass cover slips in B-27 neurobasal medium containing 0.5 mM glutamine and 25 µM glutamate (Invitrogen, Carlsbad, CA). The neuronal cells were grown under 5% CO2 in an incubator maintained at 37°C until differentiation...

Images: A–D: Uptake of fluorescein labeled Aβ40 (F-Aβ40) and Alexa Fluor® 633 labeled transferrin (AF633-Trf), clathrin-mediated endocytosis marker, in rat primary hippocampal (RPH) neurons following 30 min incubation at 37°C. (A) F-Aβ40 uptake; (B) Uptake of AF633-Trf; (C) Superimposition of images A and B; (D) Overlay of fluorescence images on the DIC image of RPH neurons. E–G: Uptake of F-Aβ40 and AF633-Trf in RPH neurons at 4°C. (E) Uptake of F-Aβ40; (F) No significant neuronal uptake of AF633-Trf at 4°C; (G) Superimposition of images D and E on the DIC image of RPH neurons; H–J: Uptake of F-Aβ40 and AF633-Trf in RPH neurons treated with 10 mM Sodium Azide and 50 mM 2-deoxy glucose, agents that are known to deplete cellular ATP. (H) Uptake of F-Aβ40; (I) No significant cellular uptake of AF633-Trf was observed; (J) Superimposition of images H and I on the DIC image of neurons.doi:10.1371/journal.pone.0004627.g010

1 comment:

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