Journal:

Article Title: Genetically altered AMPA-type glutamate receptor kinetics in interneurons disrupt long-range synchrony of gamma oscillation

doi: 10.1073/pnas.051631898

Figure Lengend Snippet: Generation of mutant mice with elevated GluR-B expression in GABAergic interneurons. (a) Schematic representation of gene segments of the wild-type GAD67 allele, the targeting vector, and the targeted GAD67 allele. Positions of EcoRI (E), NotI (N), and SalI (S) restriction sites are indicated. The PCR fragment used as probe for the Southern blot is indicated as a black bar. (Right) Southern blot analysis of tail DNA isolated from wild-type and heterozygous mice, digested with EcoRI to distinguish wild-type (7-kb) and targeted (5-kb) alleles. (b–e) GluR-B immunostaining in wild-type (b and d) and combined immunostaining and X-Gal staining in the mutant (c and e) hippocampus. Note the much higher GluR-B expression in LacZ-positive GABAergic interneurons compared with neighboring pyramidal cells in the mutant CA1 region (c) and hilus (e). This expression was never seen in the corresponding areas in control animals (b and d). (Scale bars, 100 μm.) so, Stratum oriens; sp, stratum pyramidale; sr, stratum radiatum; h, hilus. For higher magnification see Fig. 6 in the supplemental data.

Article Snippet: After X-Gal staining, the free-floating sections were incubated overnight at 4°C with a rabbit anti-GluR-B antibody (PharMingen, dilution 1:25) in 2% normal goat serum diluted in Tris-buffered saline (TBS), pH 7.4.

Techniques: Mutagenesis, Expressing, Plasmid Preparation, Southern Blot, Isolation, Immunostaining, Staining

Journal:

Article Title: Genetically altered AMPA-type glutamate receptor kinetics in interneurons disrupt long-range synchrony of gamma oscillation

doi: 10.1073/pnas.051631898

Figure Lengend Snippet: Firing properties of interneurons are altered by genetically manipulated overexpression of GluR-B. (a) Spontaneous EPSP rise time and decay time plots for stratum pyramidale fast-spiking interneurons, recorded from a membrane potential of −70 mV. Data are shown as cumulative probability for >4,000 EPSPs from five cells from five wild-type and five cells from five mutant mice. Model data show change in unitary EPSC for parameters that best fit the experimental data: “wild-type” unitary EPSC = t exp (−t/0.95) nS, “mutant” unitary EPSC = t exp(−t/1.05) nS. (Scale bars: 0.8 nS, 1 ms.) (b) Firing pattern in response to a single proximal stratum radiatum stimulation. Traces show response (from −70 mV) to increasing stimulus intensity (5–20 V) for interneurons from wild-type and mutant. Note increasing intensities generate double spikes in the cell from the mutant. (Scale bars: 50 mV, 40 ms.) Model data show voltage responses for single interneurons (holding current −0.165 nA) receiving identical EPSC onto single compartments on each of three dendrites. Each EPSC had the time course ct exp(−t/τ), where τ = 0.95 for control and 1.05 for mutant; c = 16.1, 47.4, 83.8 nS (top to bottom). Note the spike doublet in the mutant, but not in the wild type (as in experiment). (Scale bars: 50 mV, 30 ms.) (c) Pattern of pyramidal cell phasic inhibitory input (upper traces) and interneuron phasic excitatory input (lower traces) during posttetanic oscillations, in wild-type and in mutant mice. The interneuron was hyperpolarized by injection of −0.2 nA current. [Scale bars (experiment): 2 mV, 100 ms.] Model data show GABAA conductance to an e-cell (upper traces) and AMPA conductance to an i-cell. Note, in the “mutant,” the more variable amplitude of GABAA inputs, and the variable width of the AMPA inputs. [Scale bars (model): 150 nS, 100 ms.] (d) Example traces of interneuron firing patterns during gamma-frequency oscillations following paired tetanic stimulation, illustrating the increased incidence of doublet formation and the occurrence of occasional spike bursts. Model data show voltage of a selected interneuron from network simulations in the wild type and the mutant. The only difference in parameters for these simulations was in interneuron EPSC time course illustrated in a. [Scale bars (experiment and model): 20 mV, 100 ms.] Below are histograms illustrating the probability of a doublet of interval x relative to the sample mode. There are 45 doublets from wild-type data (n = three cells from three animals) and 130 doublets from mutant data (n = five cells from five animals). Model histograms were constructed by using ≈110 doublet intervals for simulations of “wild type” and “mutant” gamma oscillations, each pooled from seven interneurons.

Article Snippet: After X-Gal staining, the free-floating sections were incubated overnight at 4°C with a rabbit anti-GluR-B antibody (PharMingen, dilution 1:25) in 2% normal goat serum diluted in Tris-buffered saline (TBS), pH 7.4.

Techniques: Over Expression, Mutagenesis, Injection, Construct

Journal: PLoS ONE

Article Title: Searching for Cellular Partners of Hantaviral Nonstructural Protein NSs: Y2H Screening of Mouse cDNA Library and Analysis of Cellular Interactome

doi: 10.1371/journal.pone.0034307

Figure Lengend Snippet: Intersection of closest shared TULV and PUUV NSs -linked nodes. Primary nodes are in red, secondary nodes in pink. Secondary nodes that do not directly connect to two different primary nodes are in white. Metanodes are represented by pink diamonds.

Article Snippet: Samples were incubated 1 h at 37°C with rabbit polyclonal antibodies against TULV NSs-GST fusion protein and mouse monoclonal anti-ACBD3 antibody (Abnova, Taipei, Taiwan).

Techniques:

Journal: PLoS ONE

Article Title: Searching for Cellular Partners of Hantaviral Nonstructural Protein NSs: Y2H Screening of Mouse cDNA Library and Analysis of Cellular Interactome

doi: 10.1371/journal.pone.0034307

Figure Lengend Snippet: Primary cells were infected with TULV strain Lodz for 9 days and fixed on coverslips. Cells were stained for the viral NSs protein, which was seen as bright spots around perinuclear area (A), and ACBD3 protein (B). FRET assay: D pre , donor intensity before bleaching, D post , donor intensity after bleaching, A pre , acceptor intensity before bleaching, A post , acceptor intensity after bleaching, FRET eff , calculated efficiency of FRET.

Article Snippet: Samples were incubated 1 h at 37°C with rabbit polyclonal antibodies against TULV NSs-GST fusion protein and mouse monoclonal anti-ACBD3 antibody (Abnova, Taipei, Taiwan).

Techniques: Infection, Staining

Journal: The Journal of Biological Chemistry

Article Title: Hydrogen Sulfide Inhibits High Glucose-induced Matrix Protein Synthesis by Activating AMP-activated Protein Kinase in Renal Epithelial Cells * ^♦

doi: 10.1074/jbc.M111.278325

Figure Lengend Snippet: Expression of hydrogen sulfide-generating enzymes is reduced in the kidney in diabetes. A and B, expression of cystathionine β-synthase (CBS) and cystathionine γ-lyase (CSE) is reduced in the renal cortex of mice with type 1 diabetes (A) or type 2 (B) diabetes. Equal amounts of tissue homogenate protein were immunoblotted with antibody against CBS or cystathionine γ-lyase; loading was assessed by immunoblotting for actin (*, p < 0.05; **, p < 0.01 versus control by t test). C, immunoperoxidase staining of the kidney showed reduction in the expression of cystathionine β-synthase in type 1 diabetic mice (OVE26) when compared with non-diabetic NJ control mice.

Article Snippet: The slides were then blocked with Sniper blocking buffer (Biocare, Concord, CA) for 20 min and incubated with the rabbit polyclonal anti-cystathionine β-synthase (1:500; Abgent, San Diego, CA) overnight at 4 °C in a humidified chamber.

Techniques: Expressing, Western Blot, Immunoperoxidase Staining

Journal: The Journal of Biological Chemistry

Article Title: Hydrogen Sulfide Inhibits High Glucose-induced Matrix Protein Synthesis by Activating AMP-activated Protein Kinase in Renal Epithelial Cells * ^♦

doi: 10.1074/jbc.M111.278325

Figure Lengend Snippet: Schematic showing signaling pathways involved in hydrogen sulfide amelioration of high glucose-induced protein synthesis. CBS, cystathionine β-synthase; CSE, cystathionine γ-lyase.

Article Snippet: The slides were then blocked with Sniper blocking buffer (Biocare, Concord, CA) for 20 min and incubated with the rabbit polyclonal anti-cystathionine β-synthase (1:500; Abgent, San Diego, CA) overnight at 4 °C in a humidified chamber.

Techniques: