Journal: Animal Bioscience

Article Title: Oxidative stress impairs the meat quality of broiler by damaging mitochondrial function, affecting calcium metabolism and leading to ferroptosis

doi: 10.5713/ab.22.0019

Figure Lengend Snippet: (a, b) Effects of H 2 O 2 exposure on the contents of Ca 2+ . (c, d) Effects of H 2 O 2 exposure on the relative protein expression levels of TRPC1, PMCA, NCX, SERCA1, and MCU. The control group was the non-injected treatment. The saline group: birds were injected with physiological saline buffer (0.75%) with a dosage of 1.0 mL/kg BW. The H 2 O 2 group: birds were given an injection of 10.0% H 2 O 2 with a dosage of 1.0 mL/kg BW. Data were reported as mean±standard error (n = 6). TRPC1, transient receptor potential channel 1; PMCA, plasma membrane Ca 2+ -ATPase; NCX, sodium-calcium exchanger; SERCA1, sarcoplasmic/endoplasmic reticulum calcium ATPase 1; MCU, mitochondrial calcium uniporter; BW, body weight. a,b denoted p<0.05.

Article Snippet: The antibodies of plasma membrane Ca 2+ -ATPase (PMCA bs-4978R), transient receptor potential channel 1 (TRPC1 bs-10404R), and sodium-calcium exchanger (NCX bs-1550R) were produced by Bioss biotechnology company (Beijing, China).

Techniques: Expressing, Control, Injection, Saline, Membrane

Journal: Marine Drugs

Article Title: Selective Inhibition of Liver Cancer Cells Using Venom Peptide

doi: 10.3390/md17100587

Figure Lengend Snippet: Potential mechanism of Tv1 antitumor activity inhibits COX2 and PGE2 function. In our model of Tv1 antitumor activity in liver cancer cells, overexpression of TRP channels (TRPC6 and V6) stimulates COX-2-dependent PGE2 production via enhanced [Ca 2+ ] dynamics. Influx of Ca 2+ can occur through voltage gated (VGC), receptor operated (ROC), and store operated (SOC) calcium channels. Transient receptor potential (TRP) channels contribute to store operated calcium (SOC) channels. Ca 2+ -dependent transcription factor NFAT is activated via dephosphorylation by calcineurin, which is activated upon binding of Ca 2+ /calmodulin. Ubiquitously present transcription factor NFAT regulates COX-2 expression and further prostaglandin E2 (PGE2) release in different cancer cells and its activation occurs through Ca 2+ influx associated with TRPC1-, TRPC3-, or TRPC6-associated SOC or ROC activities. PGE2 release plays multiple roles in cancer as shown. Upon Tv1 application to liver cancer (HCC) cells the pictured downstream pathways leading to proliferation inhibition and apoptosis are encountered.

Article Snippet: Rabbit polyclonal Anti-HERG (Kv11.1)-APC109, Anti TRPV6-ACC036, Anti TRPV2-ACC039, Anti TRPC1-ACC-118 were purchased from Alomone lab, Israel.

Techniques: Activity Assay, Over Expression, De-Phosphorylation Assay, Binding Assay, Expressing, Activation Assay, Inhibition

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

Article Title: Modulation of Canonical Transient Receptor Potential Channel 1 in the Proliferation of Oligodendrocyte Precursor Cells by the Golli Products of the Myelin Basic Protein Gene

doi: 10.1523/JNEUROSCI.4424-10.2011

Figure Lengend Snippet: Sequences of primers and PCR product sizes used in the semiquantitative RT-PCR

Article Snippet: Staining with TRPC1 (1:100; Alomone Labs) was performed on live cells without permeabilization for 1h at room temperature before fixation.

Techniques: Sequencing

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

Article Title: Modulation of Canonical Transient Receptor Potential Channel 1 in the Proliferation of Oligodendrocyte Precursor Cells by the Golli Products of the Myelin Basic Protein Gene

doi: 10.1523/JNEUROSCI.4424-10.2011

Figure Lengend Snippet: Sequences of Stealth RNAi™ siRNA duplexes used for TRPC1 knockdown

Article Snippet: Staining with TRPC1 (1:100; Alomone Labs) was performed on live cells without permeabilization for 1h at room temperature before fixation.

Techniques: Sequencing

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

Article Title: Modulation of Canonical Transient Receptor Potential Channel 1 in the Proliferation of Oligodendrocyte Precursor Cells by the Golli Products of the Myelin Basic Protein Gene

doi: 10.1523/JNEUROSCI.4424-10.2011

Figure Lengend Snippet: (A) SOCC activity was examined in JOE OPCs pre-treated with thapsigargin (Tg) in the presence of zero Ca++ medium (Basal). (B) JOE OPCs were incubated with an antibody against TRPC1 (10μg/ml; 1h) before depleting their internal Ca++ stores with Tg in zero Ca++ solution (TRPC1 Ab). Compared to JOE cells in basal conditions (A), these OPCs have a significant decrease in internal Ca++ elevations when 2mM Ca++ is returned to the external solution. (C) The effect of anti-TRPC1 was totally abolished by preincubation of the antibody with the antigen peptide (+Pept). The times of addition of Tg and Ca++ containing external solutions are indicated by the horizontal bars. (D) The graphs show the average amplitude calculated from the responding cells, expressed as percentage of change of the emission intensities for each experimental condition. Values are expressed as mean ± SEM of at least four independent experiments (n>400 cells for each condition). **p<0.01 vs respective controls.

Article Snippet: Staining with TRPC1 (1:100; Alomone Labs) was performed on live cells without permeabilization for 1h at room temperature before fixation.

Techniques: Activity Assay, Incubation

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

Article Title: Modulation of Canonical Transient Receptor Potential Channel 1 in the Proliferation of Oligodendrocyte Precursor Cells by the Golli Products of the Myelin Basic Protein Gene

doi: 10.1523/JNEUROSCI.4424-10.2011

Figure Lengend Snippet: (A) RT-PCR analysis of TRPC mRNA. Equal amounts of cDNA, prepared from total RNA of control and JOE OPCs, were added for each reaction. TRPC isoform-specific PCR primers (Table I) and the semi-quantitative RT-PCR conditions used for these experiments are described in Materials and Methods. β-tubulin was used as internal standard and data are representative of three independent experiments. (B) Pure OPCs from control and JOE mice were transiently transfected with a combination of three different siRNA duplexes specific for TRPC1 (siTRPC1) and grown for 2 days in defined culture media. Semi-quantitative RT-PCR analysis of TRPC1 mRNA expression in OPCs was performed using β-tubulin as internal standard. Data from three independent experiments are summarized based on the relative spot intensities and plotted as percent of controls. (C) OPCs were grown under conditions described above. For each lane, 50μg of total protein extract were applied and Western blots performed as described under Materials and Methods. A representative Western blot for TRPC1 is shown and data from three independent experiments are summarized based on the relative spot intensities and plotted as percent of controls.

Article Snippet: Staining with TRPC1 (1:100; Alomone Labs) was performed on live cells without permeabilization for 1h at room temperature before fixation.

Techniques: Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR, Transfection, Expressing, Western Blot

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

Article Title: Modulation of Canonical Transient Receptor Potential Channel 1 in the Proliferation of Oligodendrocyte Precursor Cells by the Golli Products of the Myelin Basic Protein Gene

doi: 10.1523/JNEUROSCI.4424-10.2011

Figure Lengend Snippet: (A and B) SOCC activity was examined in JOE OPCs pre-treated with thapsigargin (Tg) in the presence of zero Ca++ medium. After 1 days in vitro (1DIV), JOE cells were transiently transfected with a combination of three different siRNA duplexes specific for TRPC1 (siTRPC1) and grown for 2 days in defined culture medium. Ca++ entry experiments were performed after 3 and 4 days of siRNA transfection (i.e., 4 and 5DIV). (C) The graphs shows the average amplitude calculated from the responding cells expressed as percentage of change of the emission intensities for each experimental condition. Values are expressed as mean ± SEM of at least four independent experiments (n>400 cells for each condition). **p<0.01 vs respective controls.

Article Snippet: Staining with TRPC1 (1:100; Alomone Labs) was performed on live cells without permeabilization for 1h at room temperature before fixation.

Techniques: Activity Assay, In Vitro, Transfection

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

Article Title: Modulation of Canonical Transient Receptor Potential Channel 1 in the Proliferation of Oligodendrocyte Precursor Cells by the Golli Products of the Myelin Basic Protein Gene

doi: 10.1523/JNEUROSCI.4424-10.2011

Figure Lengend Snippet: (A) Twenty four hours after plating, pure OPCs cultures from JOE mice were transiently transfected with a combination of three different siRNA duplexes (siTRPC1) specific for TRPC1 and grown for 2 days in vitro (DIV) in defined culture medium plus PDGF (10ng/ml) and bFGF (10ng/ml). Then the medium was changed and the cells were grown in the presence of PDGF (20ng/ml) for another 2 days (i.e., 4 and 5DIV). Twenty-four hour pulses of 10μM bromo-deoxyuridine (BrdU) were begun at 3 and 4DIV. After each BrdU pulse, cells were fixed and immunostained with anti-BrdU and anti-NG2 antibodies. (B) Microphotographs showing NG2+/BrdU+ cells at 4 and 5DIV, green: NG2 immunostaining, and red: BrdU immunostaining. Scale bar = 80μm. (C) The percentage of NG2+/BrdU+ cells in each condition was compared with respective controls. (D) The percentage of P-Histone H3+ cells in each experimental condition was measured as described in Materials and Methods. Values are expressed as mean ± SEM of four independent experiments. **p<0.01, ***p<0.001, versus respective controls.

Article Snippet: Staining with TRPC1 (1:100; Alomone Labs) was performed on live cells without permeabilization for 1h at room temperature before fixation.

Techniques: Transfection, In Vitro, Immunostaining

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

Article Title: Modulation of Canonical Transient Receptor Potential Channel 1 in the Proliferation of Oligodendrocyte Precursor Cells by the Golli Products of the Myelin Basic Protein Gene

doi: 10.1523/JNEUROSCI.4424-10.2011

Figure Lengend Snippet: (A) Mixed glial cultures were incubated in a chamber with 5% CO2 at 37°C, which was placed on the stage of a Spinning Disc Confocal Inverted Microscope. GFP-labeled OPC clones were imaged with a specific GFP filter at 6min intervals for a period of 48h beginning at 4 and 6 days in vitro (div) before the shake-off. (B) Time-lapse series of OPC∼GFP clones from JOE mice grown for 4 days on an astrocyte monolayer (4div) and incubated with an antibody against TRPC1 (TRPC1 Ab) (10μg/ml). Yellow arrowheads designate cytokinesis events. Tracking of cells between birth cytokinesis and division cytokinesis was noted with a yellow asterisk near the cell, which was generated from frame to frame. Each frame represents a single section of a time lapse video sequence. Time is denoted in hours in the bottom right corner. Scale bar = 60μm. (C and D) Estimated cell cycle times (Tcs) and percentage of mitotic OPCs were determined as described in Materials and Methods for each experimental condition. Values are expressed as mean ± SEM of four independent experiments. *p<0.05, **p<0.01, versus control.

Article Snippet: Staining with TRPC1 (1:100; Alomone Labs) was performed on live cells without permeabilization for 1h at room temperature before fixation.

Techniques: Incubation, Inverted Microscopy, Labeling, Clone Assay, In Vitro, Generated, Sequencing

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

Article Title: Modulation of Canonical Transient Receptor Potential Channel 1 in the Proliferation of Oligodendrocyte Precursor Cells by the Golli Products of the Myelin Basic Protein Gene

doi: 10.1523/JNEUROSCI.4424-10.2011

Figure Lengend Snippet: (A) Confocal microscopy analysis of JOE OPCs reveal high TRPC1 expression in certain sites along the processes and soma of the cells. Scale bar = 25μm (B) JOE OPCs were immunostained for golli (red) and TRPC1 (green). Merged images show areas of obvious co-localization and/or juxtaposition (arrows). Note that golli fluorescence is predominantly concentrated in multiple high intensity patches along the processes, and TRPC1 is found closely associated with these sites of high golli concentration (arrows). Scale bar = 15μm. (C) Time-lapse confocal microscopy of OPC process overexpressing golli-GPP during SOC influx. Peaks in local amplitude of Ca++ uptake were found at several sites along the process. The close interrelationship between golli and Ca++ influx sites can be clearly seen, with golli-GFP being closely surrounded by high levels of Fura-2 fluorescence (arrows). (D) Correlation analysis of the patterns of local peak Ca++ amplitudes and golli-GFP in the same OPC process shown in (C). Local peak Ca++ amplitudes after re-exposure to Ca++ containing medium (120s) (red line) along the process are shown compared with golli-GFP fluorescence measurement (green line) in the same cellular sections.

Article Snippet: Staining with TRPC1 (1:100; Alomone Labs) was performed on live cells without permeabilization for 1h at room temperature before fixation.

Techniques: Confocal Microscopy, Expressing, Fluorescence, Concentration Assay