Mild in vitro trauma induces rapid Glur2 endocytosis, robustly augments calcium permeability and enhances

susceptibility to secondary excitotoxic insult in cultured Purkinje cells

by Joshua D. Bell, Jinglu Ai, Yonghong Chen, and Andrew J. Baker

BrainVolume 130(10):2528-2542

September 26, 2007

© 2007 The Author(s)

Validation of Purkinje cell culture system via immunocytochemical labelling using glial fibrillary acidic protein (GFAP) (A), calbindin D-28K (B), MAP2 (C,D) as markers for astrocytes, Purkinje

cells and neurons, respectively.

Joshua D. Bell et al. Brain 2007;130:2528-2542

© 2007 The Author(s)

(A) Injury magnitude-propidium iodide (PI) uptake dose–response curve for stretch pressures ranging from 2.5 to 7.5 psi.

Joshua D. Bell et al. Brain 2007;130:2528-2542

© 2007 The Author(s)

Mild stretch robustly augments AMPA induced calcium influx as evidenced by normalized Purkinje cell Fura-2AM ratios (340 nm/380 nm) following application of 4 µM AMPA. (A) Calcium

influx is moderate, and quickly returns to baseline values in control cells.

Joshua D. Bell et al. Brain 2007;130:2528-2542

© 2007 The Author(s)

Immunocytochemical labelling of control and injured neurons for GluR2 (A & D respectively), synaptophysin (B & E respectively), and the overlay of the two (C and F respectively).

Joshua D. Bell et al. Brain 2007;130:2528-2542

© 2007 The Author(s)

(A) High-magnification images of dendritic processes of mildly injured (A1) and control (A2) Purkinje neurons.

Joshua D. Bell et al. Brain 2007;130:2528-2542

© 2007 The Author(s)

Mild injury does not increase non-specific Purkinje cell membrane permeability.

Joshua D. Bell et al. Brain 2007;130:2528-2542

© 2007 The Author(s)

(A) Combination therapy of PKCα inhibition and blockade of calcium permeable GluR1/3 containing AMPA receptors confers profound neuroprotection against delayed cell death caused

by 20 µM AMPA and 2.9 psi insult.

Joshua D. Bell et al. Brain 2007;130:2528-2542

© 2007 The Author(s)