g54  (ATCC)

Journal: Frontiers in Neuroscience

Article Title: Blockade of TRPC Channels Limits Cholinergic-Driven Hyperexcitability and Seizure Susceptibility After Traumatic Brain Injury

doi: 10.3389/fnins.2021.681144

Figure Lengend Snippet: Normalized fold change in TRPC1/4/5 mRNA (2 –ΔΔ CT ) in TBI mice compared to sham.

Article Snippet: Parietal cortex or dorsal hippocampus brain sections (−1.3 to −2.3 mm posterior to bregma) were probed with a TRPC4 mouse monoclonal antibody (1:1,000, Rockland #200-301-G54, RRID:AB_2611251 ) at 1:500 dilution and fluorescein isothiocyanate (FITC) donkey antimouse IgG (1:250, Jackson ImmunoResearch #715-095-150, RRID:AB_2340792 ).

Techniques:

Journal: Frontiers in Neuroscience

Article Title: Blockade of TRPC Channels Limits Cholinergic-Driven Hyperexcitability and Seizure Susceptibility After Traumatic Brain Injury

doi: 10.3389/fnins.2021.681144

Figure Lengend Snippet: Cell-type specific TRPC1, TRPC4, and TRPC5 channel upregulation in the hippocampus and cortex after CCI-TBI. (A,B) Representative Western immunoblots using (A) TRPC4 (∼120 kDa) and (B) TRPC5 (∼110 kDa) antibodies in sham and TBI cortex and hippocampus. Blots were normalized to β-actin protein (42 kDa) as loading control. (C,D) Summarized data for Western blot quantification of TRPC4 ( C , n = 7–13 animals per group) and TRPC5 ( D , n = 5–7 animals per group) from microdissected brain regions in mice 7 days after TBI. (E,F) Shown are summarized plots of percent difference in TRPC4 (E) and TRPC5 (F) protein between ipsilateral and contralateral hemispheres of microdissected regions from data in Panels (C,D) . All data bars represent the mean ± SEM. * p < 0.05 vs. sham of same subregion. † p < 0.05 vs. contralateral hemisphere of same subregion.

Article Snippet: Parietal cortex or dorsal hippocampus brain sections (−1.3 to −2.3 mm posterior to bregma) were probed with a TRPC4 mouse monoclonal antibody (1:1,000, Rockland #200-301-G54, RRID:AB_2611251 ) at 1:500 dilution and fluorescein isothiocyanate (FITC) donkey antimouse IgG (1:250, Jackson ImmunoResearch #715-095-150, RRID:AB_2340792 ).

Techniques: Western Blot, Control

Journal: Frontiers in Neuroscience

Article Title: Blockade of TRPC Channels Limits Cholinergic-Driven Hyperexcitability and Seizure Susceptibility After Traumatic Brain Injury

doi: 10.3389/fnins.2021.681144

Figure Lengend Snippet: Normalized TRPC4/TRPC5 protein in 7-day sham and TBI mice.

Article Snippet: Parietal cortex or dorsal hippocampus brain sections (−1.3 to −2.3 mm posterior to bregma) were probed with a TRPC4 mouse monoclonal antibody (1:1,000, Rockland #200-301-G54, RRID:AB_2611251 ) at 1:500 dilution and fluorescein isothiocyanate (FITC) donkey antimouse IgG (1:250, Jackson ImmunoResearch #715-095-150, RRID:AB_2340792 ).

Techniques:

Journal: Frontiers in Neuroscience

Article Title: Blockade of TRPC Channels Limits Cholinergic-Driven Hyperexcitability and Seizure Susceptibility After Traumatic Brain Injury

doi: 10.3389/fnins.2021.681144

Figure Lengend Snippet: Surges in neuronal activity following CCI-TBI are TRPC4/TRPC5-mediated. (A) Representative images of parietal cortex from sham and TBI TRAP mice that were administered 4-OHT at t = 12 h before procedure. Prefix “ c ” denotes contralateral, prefix “ i ” denotes ipsilateral. Red = cFos-tdTomato; blue = DAPI. (B) Representative images of hippocampal subregions from sham and TBI TRAP mice that were administered 4-OHT at t = 12 h before procedure. (C) Summarized quantification of cFos+ neuron density (neurons/0.1 mm 3 ) in sham and TBI TRAP mice activated at the time of TBI, as in Panels (A,B) . (D) Representative images taken from sham mice, TBI mice, and TBI mice also administered M084 (10 mg/kg) (TBI + M084) that were administered 4-OHT t = 7 days after procedure. (E) Summarized quantification of cFos+ neurons in sham, TBI, and TBI + M084 mice 7 days after procedure, as in Panel (D) . All data bars represent the mean ± SEM. * p < 0.05 vs. sham. # p < 0.05 vs. TBI cDG. † p < 0.05 vs. TBI of same region. Scale bars: 100 μm.

Article Snippet: Parietal cortex or dorsal hippocampus brain sections (−1.3 to −2.3 mm posterior to bregma) were probed with a TRPC4 mouse monoclonal antibody (1:1,000, Rockland #200-301-G54, RRID:AB_2611251 ) at 1:500 dilution and fluorescein isothiocyanate (FITC) donkey antimouse IgG (1:250, Jackson ImmunoResearch #715-095-150, RRID:AB_2340792 ).

Techniques: Activity Assay

Journal: Frontiers in Neuroscience

Article Title: Blockade of TRPC Channels Limits Cholinergic-Driven Hyperexcitability and Seizure Susceptibility After Traumatic Brain Injury

doi: 10.3389/fnins.2021.681144

Figure Lengend Snippet: TRPC4/TRPC5 channel activation artificially prolongs Ca 2+ influx in DGGCs after CCI-TBI. (A) Cumulative probability distribution of the peak amplitude of GCaMP6f fluorescence (ΔF/F) for each DGGC from sham (control) and iTBI slices during EA (1 μM, red) or EA + M084 (10 μM, gray) application. (B) Cumulative probability distribution of the Ca 2+ influx duration (in seconds) for each DGGC from control and iTBI slices during EA or EA + M084 application. (C,D) Histogram population distribution of control DGGC Ca 2+ influx events according to peak amplitude (C) and Ca 2+ event duration (D) . (E,F) Histogram population distribution of iTBI DGGC Ca 2+ influx events according to peak amplitude (E) and Ca 2+ event duration (F) . (G) Summarized means of peak fluorescence from data as in Panel (A) . (H) Summarized means of Ca 2+ influx duration from data as in Panel (B) . Red = EA alone, gray = EA + M084. All data bars represent the mean ± SEM. * p < 0.05 vs. EA alone from same procedure condition, † p < 0.05 vs. control of same drug condition.

Article Snippet: Parietal cortex or dorsal hippocampus brain sections (−1.3 to −2.3 mm posterior to bregma) were probed with a TRPC4 mouse monoclonal antibody (1:1,000, Rockland #200-301-G54, RRID:AB_2611251 ) at 1:500 dilution and fluorescein isothiocyanate (FITC) donkey antimouse IgG (1:250, Jackson ImmunoResearch #715-095-150, RRID:AB_2340792 ).

Techniques: Activation Assay, Fluorescence, Control

Journal: Biomedical Optics Express

Article Title: Integrated multi-wavelength microscope combining TIRFM and IRM modalities for imaging cellulases and other processive enzymes

doi: 10.1364/BOE.423798

Figure Lengend Snippet: Microscope schematic. Illumination is provided by six lasers at left that are each expanded and collimated by a pair of lenses (Lf and Le) and a pinhole (Ph), Irises (Ir) are used to adjust the beam size after expansion. The beams are combined and collimated by a series of dichroic mirrors (DM1-DM5). The 785 nm beam used for auto-focus is reflected off of a broadband mirror (BP). This combined illumination beam is used for dichroic-based illumination (1) or micromirror-based illumination (2), depending on the position of flip mirrors M1 and M2. In the dichroic pathway, the position of mirrors M3 and M4 are adjusted to switch between TIRFM and epi-fluorescence modes. IRM illumination is supplied by an LED that is reflected off a dichroic mirror below the objective. Images pass through a filter wheel (FW) and dual-view system (DV) before being captured by the camera. Auto-focus is achived using a quadrant photodiode downstream of a bandpass filter (F1) and lens (L2). At right is a detailed view of the objective and associated components, showing micromirrors (MM1 and MM2) for TIR excitation in the back of the objective, and the dichroic mirror (DM6) used for both IRM and dichroic-based TIRFM or epi-fluorescence illumination. Note that only the lenses (L1 and L2) used for micromirror TIR excitation are shown in figure at left; lenses for all beam paths are shown at right. See text for full description of components.

Article Snippet: Following total internal reflection at the glass-water interface of the sample, the beam returns through the objective and is reflected by a second micromirror (MM2, G54-092, Edmund Optics; USA), passes through a focusing lens (L2, f = 150 mm) and a bandpass fiter (F1, et775-50x, Chroma; USA), and arrives at a quadrant photodiode (QPD; TIRF Lock, Mad City Labs; USA).

Techniques: Microscopy, Fluorescence

Journal: Biomedical Optics Express

Article Title: Integrated multi-wavelength microscope combining TIRFM and IRM modalities for imaging cellulases and other processive enzymes

doi: 10.1364/BOE.423798

Figure Lengend Snippet: Simultaneous imaging of cellulases and cellulose. a) IRM images of label-free cellulose (left) and TIRFM images of S4B labeled cellulose (right) in the same region. b) Intensity profile comparison of IRM (blue line) and TIRFM (red line). The IRM image intensity was inverted and scaled to show overlap with fluorescence. c) Composite image of immobilized cellulose imaged by IRM (black fibers on the gray background) with Qdot525-labeled-Cel7a (Green) and Qdot655-labeled Cel6a (Magenta) imaged by two-color micromirror TIRFM.

Article Snippet: Following total internal reflection at the glass-water interface of the sample, the beam returns through the objective and is reflected by a second micromirror (MM2, G54-092, Edmund Optics; USA), passes through a focusing lens (L2, f = 150 mm) and a bandpass fiter (F1, et775-50x, Chroma; USA), and arrives at a quadrant photodiode (QPD; TIRF Lock, Mad City Labs; USA).

Techniques: Imaging, Labeling, Comparison, Fluorescence

Journal: Studies in Mycology

Article Title: Aspergillus fumigatus and aspergillosis: From basics to clinics

doi: 10.1016/j.simyco.2021.100115

Figure Lengend Snippet: Studies using in-house and commercial methodologies for the direct detection of azole resistance in A. fumigatus from clinical samples.

Article Snippet: Pyrosequencing , In-house , G54 , Blood (56) , 3.6 % , , 3.6 % WT , Trama et al. 2005 .

Techniques: Amplification, Polymerase Chain Reaction, Sequencing, Real-time Polymerase Chain Reaction, Clinical Proteomics, Nested PCR

Journal: Studies in Mycology

Article Title: Aspergillus fumigatus and aspergillosis: From basics to clinics

doi: 10.1016/j.simyco.2021.100115

Figure Lengend Snippet: Consequences of antifungal resistance of Aspergillus species in murine models and Galleria .

Article Snippet: Pyrosequencing , In-house , G54 , Blood (56) , 3.6 % , , 3.6 % WT , Trama et al. 2005 .

Techniques: Infection, In Vitro, In Vivo