Journal: Journal of Cerebral Blood Flow & Metabolism

Article Title: Cerebrospinal fluid high mobility group box 1 is associated with neuronal death in subarachnoid hemorrhage

doi: 10.1177/0271678X16629484

Figure Lengend Snippet: Administration of sRAGE protects rat brain against SAH-induced cell death and DNA damage. Intrathecal injection of recombinant sRAGE (0.l g/rat) at the time of the SAH procedure significantly reduced the number of TUNEL-positive brain cells (a and b) and 8OHDG expression (c and d) compared to controls at both 24 and 48 h after SAH injury. **p < 0.01 versus control group at 24 h. ***p < 0.001 versus control group at 48 h.

Article Snippet: After blocking endogenous peroxidase with 0.3% hydrogen peroxidase in methanol for 30 min, the slides were washed and then incubated with anti-8OHdG monoclonal antibody (diluted 1:100; NOF corporation, Japan) overnight at 4°C and exposed to goat anti-mouse immunoglobulin conjugated to peroxidase-labeled dextran polymer for 30 min, followed by staining with 3,3′-diaminobenzidine tetrahydrochloride.

Techniques: Injection, Recombinant, TUNEL Assay, Expressing, Control

Journal: The Journal of Neuroscience

Article Title: Reverse of Age-Dependent Memory Impairment and Mitochondrial DNA Damage in Microglia by an Overexpression of Human Mitochondrial Transcription Factor A in Mice

doi: 10.1523/JNEUROSCI.1957-08.2008

Figure Lengend Snippet: Protein levels of human TFAM overexpressed by the tetracycline-off system in HeLa cells. A, Immunoblot analysis of human TFAM expressed in HeLa cells cultured with and without DC for 24 h. B, The mean protein level of human TFAM in HeLa cells cultured with and without DC for 6 and 24 h. The mean relative immunoreactivity of each protein band was determined using the level of actin as an internal control. Each column and bar represent the mean and SEM of three experiments, respectively.

Article Snippet: Floating coronal sections (10 μm thick) of the hippocampus were prepared by a cryostat and stained with anti-human TFAM, anti-8-oxo-deoxyguanosine (8-oxo-dG) (NOF Corporation), anti-4-hydroxy-2-nonenal (HNE) (Alpha Diagnostic), and anti-interleukin-1β (IL-1β) (Santa Cruz Biotechnology) for 3 d at 4°C.

Techniques: Western Blot, Cell Culture, Control

Journal: The Journal of Neuroscience

Article Title: Reverse of Age-Dependent Memory Impairment and Mitochondrial DNA Damage in Microglia by an Overexpression of Human Mitochondrial Transcription Factor A in Mice

doi: 10.1523/JNEUROSCI.1957-08.2008

Figure Lengend Snippet: Effects of an overexpression of TFAM by the tetracycline-off system on rotenone-induced intracellular ROS generation and NF-κB nuclear translocation in HeLa cells. A, CLMS images of rotenone-induced intracellular ROS generation measured using a ROS-sensitive dye (green), DHE, in HeLa cells cultured with or without DC. Scale bar, 20 μm. B, The mean DHE oxidation levels in HeLa cells cultured with or without DC after treatment with EtOH or rotenone. Each column and bar represent the mean ± SEM of six experiments. The asterisks indicate a significant difference between two groups (*p < 0.05; **p < 0.01; ***p < 0.001). C, NF-κB (green) and propidium iodide (PI)-stained nuclei (red) in HeLa cells cultured with or without DC after the treatment with EtOH or rotenone. The arrowheads show HeLa cells with the nuclear translocated NF-κB. Scale bar, 50 μm. D, The mean percentage of the NF-κB nuclear translocation in HeLa cells cultured with or without DC after the treatment with EtOH or rotenone. Each column and bar represent the mean ± SEM of nine experiments. The asterisks indicate significant differences between two groups (*p < 0.05; ***p < 0.001). Rot, Rotenone.

Article Snippet: Floating coronal sections (10 μm thick) of the hippocampus were prepared by a cryostat and stained with anti-human TFAM, anti-8-oxo-deoxyguanosine (8-oxo-dG) (NOF Corporation), anti-4-hydroxy-2-nonenal (HNE) (Alpha Diagnostic), and anti-interleukin-1β (IL-1β) (Santa Cruz Biotechnology) for 3 d at 4°C.

Techniques: Over Expression, Translocation Assay, Cell Culture, Staining

Journal: The Journal of Neuroscience

Article Title: Reverse of Age-Dependent Memory Impairment and Mitochondrial DNA Damage in Microglia by an Overexpression of Human Mitochondrial Transcription Factor A in Mice

doi: 10.1523/JNEUROSCI.1957-08.2008

Figure Lengend Snippet: The effects of TFAM overexpression on age-dependent increased oxidative stress in the brain. A, Expression of TFAM in the brain. Immunoblotting was performed using specific antibodies against human TFAM (h-TFAM) and mouse Tfam (m-Tfam) in soluble brain extracts prepared from the young WT and TG mice. B, Immunohistochemical staining for H-TFAM in the CA1 hippocampal subfield of aged WT and TG mice. Scale bar, 50 μm. C, The mean lipid peroxidation levels measured using a biochemical assay for TBARS in the brain tissues prepared from the WT and TG mice of both age groups. Each column and bar represent the mean ± SEM of five experiments. An asterisk indicates a significant difference versus the young WT mice (*p < 0.05). A dagger indicates a significant difference versus the aged WT mice (†p < 0.05). D, The mean activity of mitochondrial respiratory enzymes, complexes I to IV, in brain tissues prepared from the WT and TG mice of both age groups. Each column and bar represent the mean ± SEM of five experiments. The asterisks indicate significant differences versus the young WT mice (*p < 0.05). The daggers indicate significant differences versus the aged WT mice (†p < 0.05).

Article Snippet: Floating coronal sections (10 μm thick) of the hippocampus were prepared by a cryostat and stained with anti-human TFAM, anti-8-oxo-deoxyguanosine (8-oxo-dG) (NOF Corporation), anti-4-hydroxy-2-nonenal (HNE) (Alpha Diagnostic), and anti-interleukin-1β (IL-1β) (Santa Cruz Biotechnology) for 3 d at 4°C.

Techniques: Over Expression, Expressing, Western Blot, Immunohistochemical staining, Staining, Activity Assay

Journal: The Journal of Neuroscience

Article Title: Reverse of Age-Dependent Memory Impairment and Mitochondrial DNA Damage in Microglia by an Overexpression of Human Mitochondrial Transcription Factor A in Mice

doi: 10.1523/JNEUROSCI.1957-08.2008

Figure Lengend Snippet: Effects of TFAM overexpression on basal synaptic transmission in the hippocampus. A, Superimposed traces of field EPSPs evoked by different stimulus intensities (5–40 V) from the CA1 subfield of hippocampal slices prepared from the aged WT and TG mice. B, The input–output relationship for the mean field EPSP slope plotted against stimulus intensity in the Schaffer collateral–CA1 synapses. Open circles, WT mice; filled circles, TG mice. The point and bar represent the mean and SEM of eight slices from eight animals in the aged WT group, and seven slices from three animals in the aged TG group. No significant difference was observed between the WT and TG slices. C, Superimposed traces of field EPSPs evoked by paired pulse stimulation of the stratum radiatum with five different interspike intervals in the CA1 subfield of hippocampal slices prepared from the aged WT and TG mice. D, The plots of the relative amplitude of the second stimulation to the first responses against various interstimulus intervals. Open circles, WT mice; filled circles, TG mice. The point and bar represent the mean and SEM of six to eight slices from six animals in the aged WT mice, and seven slices from seven animals in aged TG mice. No significant difference was observed between WT and TG slices.

Article Snippet: Floating coronal sections (10 μm thick) of the hippocampus were prepared by a cryostat and stained with anti-human TFAM, anti-8-oxo-deoxyguanosine (8-oxo-dG) (NOF Corporation), anti-4-hydroxy-2-nonenal (HNE) (Alpha Diagnostic), and anti-interleukin-1β (IL-1β) (Santa Cruz Biotechnology) for 3 d at 4°C.

Techniques: Over Expression, Transmission Assay

Journal: The Journal of Neuroscience

Article Title: Reverse of Age-Dependent Memory Impairment and Mitochondrial DNA Damage in Microglia by an Overexpression of Human Mitochondrial Transcription Factor A in Mice

doi: 10.1523/JNEUROSCI.1957-08.2008

Figure Lengend Snippet: Increased expression of IL-1β in the hippocampus after the systemic injection of LPS and the inhibitory effect of the TFAM overexpression. A, Immunoblot analysis of IL-1β in the whole brain of the aged WT and TG mice subjected to intraperitoneal injection of saline (S) or LPS (0.33 mg/kg) 4 h earlier. B, The mean protein level of IL-1β. The mean relative immunoreactivity was determined using the level of actin as an internal control. Each column and bar represent the mean and SEM of three experiments, respectively. The asterisks indicate significant differences between the values (*p < 0.05). C, Immunofluorescent CLSM images for IL-1β (green) with the microglial marker Iba1 (red; top row), and 8-oxo-dG (green) with the mitochondrial marker Cyt b (red; bottom row) in the hippocampal CA1 subfield of the aged WT, the aged TG, and the LPS-treated aged TG mice. Scale bars: top row, 10 μm; bottom row, 5 μm. D, The mean immunofluorescence intensity of IL-1β/cell in the hippocampal CA1 subfield of the WT, the TG, and the LPS-treated TG mice of the aged group. Each column and bar represent the mean ± SEM of nine sections from three animals. The asterisks indicate significant differences between the values (*p < 0.05). E, The mean immunofluorescence intensity of 8-oxo-dG/mitochondria in the hippocampal CA1 subfield of the WT, the TG, and the LPS-treated TG mice of the aged group. Each column and bar represent the mean ± SEM of seven sections from two animals. The asterisks indicate significant differences between the values (***p < 0.0001; *p < 0.05).

Article Snippet: Floating coronal sections (10 μm thick) of the hippocampus were prepared by a cryostat and stained with anti-human TFAM, anti-8-oxo-deoxyguanosine (8-oxo-dG) (NOF Corporation), anti-4-hydroxy-2-nonenal (HNE) (Alpha Diagnostic), and anti-interleukin-1β (IL-1β) (Santa Cruz Biotechnology) for 3 d at 4°C.

Techniques: Expressing, Injection, Over Expression, Western Blot, Saline, Control, Marker, Immunofluorescence