The Journal of Neuroscience, May 31, 2006, ():
Different Conformations of Amyloid 
Induce Neurotoxicity by Distinct Mechanisms in Human Cortical Neurons
J. Neurosci. Deshpande et al.
26: 6011
Supplemental data
Files in this Data Supplement:
- supplemental material
-
Supplemental Figure 1. HCNs establish numerous synaptic contacts.
Triple immunofluorescence with antibodies against PSD-95 (post-synaptic marker, red fluorescence), synapthophysin (pre-synaptic marker, blue fluorescence), and tau (axonal marker; green fluorescence). The colocalization of PSD-95 and synaptophysin (white/yellow fluorescence) denotes the presence of synaptic contacts. HCN were cultured for 18 days, fixed, and processed as described in the methods section.
- supplemental material
-
Supplemental Figure 2. Treatment with AβO induces chronic mitochondrial alterations in HCN.
HCN cultures were treated with AßO at the indicated concentrations and mitochondrial membrane potential (MMP), ATP levels and oxidoreductase activity were quantified at the indicated time points. Values are expressed as mean ± SEM by unpaired Student's t test. *p< 0.05; **p<0.01. Similar results were obtained in 3 independent experiments.
- supplemental material
-
Supplemental Figure 3. Time line and proposed toxic model for Aß soluble species.
A) Time line of the progression of AßO- and ADDLs-induced neurotoxicity: I) synaptic localization of AßO and ADDLs; II) reduced mitochondrial membrane potential (MMP) and ATP levels; III) reduced mitochondrial oxidoreductase activity (MTS); IV) cyt C and AIF translocation from the mitochondrial matrix to the cytosol; V) increase caspase activity; VI) increased LDH levels in culture medium; massive neuritic retraction and disintegration, and nuclear condensation. The time line for AßO is 24 hr. ADDLs are already detected at synaptic sites by 1 hr, but progression of similar alterations takes 96 hr. Aßf (20 µM) requires 10 days to induce generalized neuronal dystrophy and modest cell death. Lower concentrations (nM range) of AßO and ADDLs lead to chronic mitochondrial dysfunction and minimal cell death over time. B) The alterations included in the time line (I to VI) are illustrated taking place at a synapse. AßO or ADDLs (red circles) are recruited to synaptic sites, where they associate with metal ions (Cu+2 and Zn+2) or bind post-synaptic receptors (I). The pore/channel-forming activity of Aß soluble species causes massive calcium influx, general destabilization of ion homeostasis and mitochondrial alterations (II: reduced MMP and ATP; III: reduced MTS), leading to the opening of the mitochondrial transition pore (MTP) and cyt C and AIF translocation (IV), caspase activation (V) and cell death evidenced by LDH release (VI). PSD: post-synaptic density
- supplemental material
-
Supplemental Movie: Time-lapse recording of 18 day HCNs treated with 5 µM AßO during 24 hr. The first and last images of this sequence correspond to Figures 2D and E respectively. Neuritic retraction is indicated by red arrows, neuritic beading and disintegration is indicated by blue arrows, and nuclear condensation is indicated by yellow marks.
