Journal: PLoS ONE

Article Title: Tg2576 Cortical Neurons That Express Human Ab Are Susceptible to Extracellular Aβ-Induced, K + Efflux Dependent Neurodegeneration

doi: 10.1371/journal.pone.0019026

Figure Lengend Snippet: Tau immunolabelling of the axonal cytoskeleton demonstrated that axonal morphology was similar between wildtype (A) and Tg2576 (B) cortical neurons over 7–14 DIV. Six daily 1 µM Aβ 1-40 treatments of 7DIV wildtype neurons had no discernible effect upon axonal morphology (C). However, substantial changes in tau-labelling were observed in Aβ 1-40 treated Tg2576 neurons (D); including increased intensity of tau immunostaining after 24 hours, followed by blebbing and axonal fragmentation which worsened after four days of treatment (D). Furthermore, after four consecutive days of 1 µM Aβ 1-40 treatment a number of axonal swellings, with dense accumulations of hyperphosphorylated tau, were observed in Tg2576 neuron cultures (E). scale bars = 30 µm (A–D), 15 µm (E).

Article Snippet: For immunocytochemistry, rabbit anti-tau (1∶5000; DAKO), mouse anti-Aβ (6E10; 1∶1000; DAKO) and mouse anti-hyperphosphorylated tau (AT-8; 1∶1000; Chemicon) antibodies were applied, and detected with appropriate Alexa-Fluor-488 or -594 conjugated secondary antibodies at a 1∶1000 concentration (Molecular Probes).

Techniques: Immunostaining

Journal: Molecular Neurodegeneration

Article Title: Intracellular amyloid formation in muscle cells of Aβ-transgenic Caenorhabditis elegans : determinants and physiological role in copper detoxification

doi: 10.1186/1750-1326-4-2

Figure Lengend Snippet: Transgenic C. elegans strains express the Aβ wild type and its variants Arctic and NIC . A . Amino acidic sequences of the Aβ 1–42 peptide (wild type) and its variants Arctic and NIC expressed in the transgenic C. elegans strains used in this study. B . PCR results show the presence of the DNAs that encode the different peptides in the transgenic strains. CL2120 is the strain that express Aβ wt and CL2122 is the control strain that do not express the peptide. NIC corresponds to transgenic line NIC2 and Arctic corresponds to transgenic line Arctic 6. unc-119 is the strain used to construct the transgenic lines carrying the NIC and Arctic variants. C . Western blot analysis using the anti-Aβ antibody 6E10 showing the expression of Aβ in the CL2120 (wild type Aβ) strain and in two NIC transgenic lines (NIC 1 and NIC 2). The lane marked as "no Aβ " correspond to strain CL2122. D . Western blot analysis using the same antibody as in C showing the expression of Aβ in strain CL2120 (wild type Aβ) and in four Arctic transgenic lines (Arctic 13, Arctic 6, Arctic 37, Arctic 5D). The lane marked as "no Aβ " correspond to strain CL2122. α-Tubulin was used as loading control in C and D.

Article Snippet: We used the anti-Aβ monoclonal antibody 6E10 (Chemicon) at 1:1,000 dilution or anti-α-tubulin monoclonal antibody (Sigma) at 1:5,000 dilution overnight at 4°C.

Techniques: Transgenic Assay, Construct, Western Blot, Expressing

Journal: The Journal of neuroscience : the official journal of the Society for Neuroscience

Article Title: Synergistic effects of long-term antioxidant diet and behavioral enrichment on beta-amyloid load and non-amyloidogenic processing in aged canines

doi: 10.1523/JNEUROSCI.6194-09.2010

Figure Lengend Snippet: (A) Representative samples from the canine parietal cortex show the anti-oligomer A11 antibody detecting the Aβ oligomeric protein at 56kDa. (B) Lower levels and reduced variability of the 56kDa aggregate are seen in the combination treatment group EA despite a large amount of variability within the groups. (CC=control environment/control diet, EC=behavioral enrichment/control diet, CA=control environment/antioxidant diet, EA=behavioral enrichment/antioxidant diet, IP=immunoprecipitated canine sample with anti-Aβ1-16 6E10 antibody, numbers on left in A=kilodalton marker, kDa=kilodalton, OD=optical density; GAPDH=glyceraldehyde 3-phosphate dehydrogenase)

Article Snippet: The antibodies used in Western blots were β-actin (rabbit, 1:5000, Abcam Inc., Cambridge, MA), GAPDH (rabbit, 1:10,000; Chemicon, Temecula, CA), the N-terminal anti-Aβ antibody 6E10 for APP (mouse, 1:5000, Signet Labs. Inc., Dedham, MA), the C-terminal anti-Aβ antibody CT20 for full length APP and CTF alpha (CTFα) and beta (CTFβ) (rabbit, 1:2000, raised against the C-terminal 20 amino acids of APP), antibodies to clearance enzymes IDE (mouse, 1:250, Covance) and neprilysin (rabbit, 1:50, Abcam Inc., Cambridge, MA), and an antibody to the precursor and mature form of the alpha secretase (αSEC) enzyme TACE/ADAM17 (rabbit, 1:2000, Chemicon, Temecula, CA) or ADAM10 (rabbit, 1:500, Chemicon, Temecula, CA).

Techniques: Immunoprecipitation, Marker