Journal: The Journal of Neuroscience

Article Title: Interaction of Survival and Death Signaling in Basal Forebrain Neurons: Roles of Neurotrophins and Proneurotrophins

doi: 10.1523/JNEUROSCI.1560-06.2006

Figure Lengend Snippet: Pro-NGF elicits loss of BF neurons in culture. BF neurons were grown for 5 d and treated overnight with the following: vehicle (imidazole), mature NGF (100 ng/ml), pro-NGF (1 ng/ml), pro-NGF (1 ng/ml) in the presence of blocking antibodies to NGF (Sigma; which also block pro-NGF; diluted in the medium 1:500) or p75 (9651 (Huber and Chao, 1995; Friedman, 2000); diluted 1:500 in the medium), or the antibodies alone. Data are expressed as mean percentage of control ± SEM in triplicate cultures from five independent experiments (n = 15). The asterisk indicates a value different from control at p < 0.05. Veh, Vehicle; mNGF, mature NGF.

Article Snippet: Primary antisera were directed against the following: p75 [192 IgG; Chemicon; 1:1000 dilution; or 9651 ( Huber and Chao, 1995 ; Friedman, 2000 ); 1:1000 dilution]; the intracellular domain of TrkB (anti-TrkB in ) [1:2000 ( Allendoerfer et al., 1994 ); or Promega; 1:1000], indicating the presence of the full-length TrkB receptors; anti-phospho-Trk, anti-phospho-Akt, or anti-cleaved caspase-3 (Cell Signaling Technologies), and anti-sortilin (anti-neurotensin receptor 3; Alpha Diagnostics; 1:1000).

Techniques: Blocking Assay

Journal: The Journal of Neuroscience

Article Title: Interaction of Survival and Death Signaling in Basal Forebrain Neurons: Roles of Neurotrophins and Proneurotrophins

doi: 10.1523/JNEUROSCI.1560-06.2006

Figure Lengend Snippet: Sortilin is required for proneurotrophin-induced death of BF neurons. A, Pro-NGF and pro-BDNF elicit a loss of BF neurons that can be prevented by either anti-p75NTR or anti-sortilin. BF neurons were cultured for 5 d and treated as indicated overnight. Pro-NGF and pro-BDNF were used at 1 ng/ml, and the antibodies were diluted 1:500 in the media. The antibodies alone had no effect on neuronal survival. B, Neurotensin blocks pro-NGF and pro-BDNF-induced neuronal loss in a dose-dependent manner. BF neurons were cultured for 5 d and treated overnight with pro-NGF (1 ng/ml) or pro-BDNF (1 ng/ml) in the presence or absence of 4 or 40 μm neurotensin. For A and B, data are expressed as mean percentage of control ± SEM in triplicate cultures from three independent experiments (n = 9). The asterisks indicate values different from control at p < 0.05. C, Immunostaining demonstrates colocalization of sortilin with p75 on cultured BF neurons. The arrows indicate p75/sortilin-positive apoptotic neurons, and the arrowheads show healthy p75/sortilin-positive neurons. Scale bar, 10 μm.

Article Snippet: Primary antisera were directed against the following: p75 [192 IgG; Chemicon; 1:1000 dilution; or 9651 ( Huber and Chao, 1995 ; Friedman, 2000 ); 1:1000 dilution]; the intracellular domain of TrkB (anti-TrkB in ) [1:2000 ( Allendoerfer et al., 1994 ); or Promega; 1:1000], indicating the presence of the full-length TrkB receptors; anti-phospho-Trk, anti-phospho-Akt, or anti-cleaved caspase-3 (Cell Signaling Technologies), and anti-sortilin (anti-neurotensin receptor 3; Alpha Diagnostics; 1:1000).

Techniques: Cell Culture, Immunostaining

Journal: The Journal of Neuroscience

Article Title: Interaction of Survival and Death Signaling in Basal Forebrain Neurons: Roles of Neurotrophins and Proneurotrophins

doi: 10.1523/JNEUROSCI.1560-06.2006

Figure Lengend Snippet: Kainic acid induced fewer dying neurons in the basal forebrain of p75−/− mice than wild-type mice. A, In wild-type mice, kainic acid induced Fluoro-Jade B (FJ) labeling, indicative of dying neurons, in p75+ cells labeled with the 9651 antibody. B, Fluoro-Jade B labeling demonstrated that kainic acid induced death of fewer neurons after 1 d in the p75−/− mice compared with wild-type mice in both the medial septal area (a, b) and diagonal band (c, d). Scale bar, 100 μm.

Article Snippet: Primary antisera were directed against the following: p75 [192 IgG; Chemicon; 1:1000 dilution; or 9651 ( Huber and Chao, 1995 ; Friedman, 2000 ); 1:1000 dilution]; the intracellular domain of TrkB (anti-TrkB in ) [1:2000 ( Allendoerfer et al., 1994 ); or Promega; 1:1000], indicating the presence of the full-length TrkB receptors; anti-phospho-Trk, anti-phospho-Akt, or anti-cleaved caspase-3 (Cell Signaling Technologies), and anti-sortilin (anti-neurotensin receptor 3; Alpha Diagnostics; 1:1000).

Techniques: Labeling

Journal: The Journal of Neuroscience

Article Title: Interaction of Survival and Death Signaling in Basal Forebrain Neurons: Roles of Neurotrophins and Proneurotrophins

doi: 10.1523/JNEUROSCI.1560-06.2006

Figure Lengend Snippet: Pro-NGF induces apoptosis of TrkB+ neurons

Article Snippet: Primary antisera were directed against the following: p75 [192 IgG; Chemicon; 1:1000 dilution; or 9651 ( Huber and Chao, 1995 ; Friedman, 2000 ); 1:1000 dilution]; the intracellular domain of TrkB (anti-TrkB in ) [1:2000 ( Allendoerfer et al., 1994 ); or Promega; 1:1000], indicating the presence of the full-length TrkB receptors; anti-phospho-Trk, anti-phospho-Akt, or anti-cleaved caspase-3 (Cell Signaling Technologies), and anti-sortilin (anti-neurotensin receptor 3; Alpha Diagnostics; 1:1000).

Techniques: