Journal: Journal of Cellular and Molecular Medicine

Article Title: Increased rhythmicity in hypertensive arterial smooth muscle is linked to transient receptor potential canonical channels

doi: 10.1111/j.1582-4934.2009.00890.x

Figure Lengend Snippet: Expression of TRPC1, TRPC3 and TRPC5 channels in mesenteric arterioles from SHR. Representative immunoblottings (A) and summary data (B) of TRPC1, TRPC3, TRPC4, TRPC5,TRPC6 channels and TRPC3 antibody with its respective antigenic peptide the protein expression in mesenteric arterioles from WKY (open bars) and SHR (filled bars). Data are mean ± S.E.M. of n = 4 independent experiments. * P < 0.05 or ** P < 0.01 by two-tailed Student’s t-test. (C) Light microscopy of mesenteric arterioles from SHR (upper panels) and immunohistochemistry (lower panels) of TRPC1, TRPC3 and TRPC5 protein expression in mesenteric arterioles from SHR. (D) Light microscopy of mesenteric arterioles from WKY (upper panels) and immunohistochemistry (lower panels) of TRPC1, TRPC3 and TRPC5 protein expression in mesenteric arterioles from WKY. Immunohistochemistry using specific antibodies labelled with green shows TRPC1, TRPC3 and TRPC5 protein expression in mesenteric arterioles. Control indicates immunohistochemistry after pre-incubation of the primary TRPC3 antibody with antigenic peptide for 12 hrs at 4°C. Bar denotes 200 μm. (43.0 ± 15%; P = n.s.; n = 6, ). In the presence of anti-TRPC5 antibodies the norepinephrine-induced vasomotion was not significantly affected (45.0 ± 11%; P = n.s. ). In the presence of anti-TRPC1 plus anti-TRPC3 antibodies the norepinephrine-induced vasomotion was significantly reduced to 29.0 ± 8% ( P < 0.05; n = 6, ). Furthermore, pre-incubation of mesenteric arterioles with anti-β-actin antibodies did not significantly affect norepinephrine-induced vasomotion when compared to control conditions (59.8 ± 15%, P = n.s.; n = 6, ). Incorporation of TRPC antibodies into mesenteric arteries by pinocytosis was facilitated by changing the bathing solution to a hypotonic solution according to recent literature .

Article Snippet: The membranes were incubated with primary rabbit antibodies against TRPC1, TRPC3, TRPC4, TRPC5 and TRPC6 (1:200; Alomone Labs), followed by incubation with goat anti-rabbit horseradish peroxidase-conjugated secondary antibody.

Techniques: Expressing, Two Tailed Test, Light Microscopy, Immunohistochemistry, Incubation

Journal: Journal of Biological Chemistry

Article Title: Inositol 1,4,5-Trisphosphate (IP3) Receptor Up-regulation in Hypertension Is Associated with Sensitization of Ca2+ Release and Vascular Smooth Muscle Contractility

doi: 10.1074/jbc.m113.496802

Figure Lengend Snippet: FIGURE 2. Screening of Ca2-handling proteins reveals coupled up-regu- lationofthevascularIP3RandLTCCinexperimentalmodelsofhyperten- sion. A, immunoblot analysis of IP3R1, 1C, TRPC1, TRPC4, Orai1, and STIM1 in lysates from mouse MA. MA lysate pooled from three SAL or AHT mice was added to each lane. Increased expression levels of IP3R1, 1C, and TRPC4 are detected in MA of AHT mice, whereas levels of TRPC1, Orai1, and STIM1 were similar. B, agarose gel analysis of products obtained after PCRs for IP3R1 (274 bp), 1C (185 bp), and TRPC4 (226 bp) using specific primers. -Actin (152 bp) and vWF (178 bp) were used as VSM and endothelial cell markers, respec- tively.IP3R,1C,and-actinweredetectedbothinintactMAandisolatedVSM cells as expected. In contrast, TRPC4 and vWF were detected in arteries but not isolated VSM cells, implying expression only in endothelium. C, immuno- blots comparing the expression of IP3R, 1C, PMCA, and -actin (as a loading control) between MA from normotensive Wistar Kyoto rats (WKY) and SHR and between MA from sham-operated and aortic-banded (Band) hyperten- sive rats. In each case, lysates were pooled from arteries of three or four rats. Only the IP3R and 1C proteins are up-regulated in MA from hypertensive SHR and aortic-banded rats. Blots are representative of three experiments.

Article Snippet: Forty g of protein was loaded per well, and expression of Ca2 -handling proteins was detected using specific antibodies against IP3R1 (1:1000; Millipore), 1C (1:1000; Millipore), TRPC1 (1:200; Santa Cruz Biotechnology), TRPC4 (1:200;NovusBiologicals), STIM1 (1:1000; Cell SignalingTechnology), Orai1 (1:1000; Sigma), and PMCA1 (1:500; Santa Cruz Biotechnology).

Techniques: Western Blot, Expressing, Agarose Gel Electrophoresis, Isolation, 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: FIGURE 2 | 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: FIGURE 3 | 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 mm3) 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: FIGURE 5 | TRPC4/TRPC5 channel activation artificially prolongs Ca2+ influx in DGGCs after CCI-TBI. (A) Cumulative probability distribution of the peak amplitude of GCaMP6f fluorescence (1F/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 Ca2+ 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 Ca2+ influx events according to peak amplitude (C) and Ca2+ event duration (D). (E,F) Histogram population distribution of iTBI DGGC Ca2+ influx events according to peak amplitude (E) and Ca2+ event duration (F). (G) Summarized means of peak fluorescence from data as in Panel (A). (H) Summarized means of Ca2+ 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, Control

Journal: British Journal of Pharmacology

Article Title: Functional expression and pharmacological modulation of TRPM3 in human sensory neurons

doi: 10.1111/bph.14994

Figure Lengend Snippet: TaqMan™ gene expression assays used in this study

Article Snippet: Sample threshold cycles were analysed relative to housekeeping gene HPRT as endogenous control (ΔC t ), and data are shown as 2 −ΔCt (mean ± SEM). table ft1 table-wrap mode="anchored" t5 Table 1 caption a7 Gene Assay TRPA1 Hs00175798_m1 TRPM3 Hs00257553_m1 TRPM8 Hs01066596_m1 TRPV1 Hs00218912_m1 SCN9A (Nav1.7) Hs01076699_m1 SCN10A (Nav1.8) Hs01045137_m1 GAPDH Hs99999905_m1 OPRM1 (μ‐opioid receptor) Hs01053957_m1 HPRT1 Hs01003267_m1 Isl‐1 Hs00158126_m1 NTRK1 (TrkA) Hs01021011_m1 NTRK2 (TrkB) Hs00178811_m1 PRDM12 Hs00964106_m1 NANOG Hs02387400_g1 CALCA (CGRP) Hs01100741_m1 POU5F1B Hs04995079_g1 RUNX1 Hs01021970_m1 TAC1 Hs00243225_m1 TRPV2 Hs00901640_m1 TRPV3 Hs00376854_m1 TRPV4 Hs01099348_m1 TRPV5 Hs00219765_m1 TRPV6 Hs00367960_m1 TRPM1 Hs00170127_m1 TRPM2 Hs01066071_m1 TRPM4 Hs00214167_m1 TRPM5 Hs00175822_m1 TRPM6 Hs00214306_m1 TRPM7 Hs00292383_m1 TRPC1 Hs00608195_m1 TRPC2 Hs03453918_m1 TRPC3 Hs00162985_m1 TRPC4 Hs01077392_m1 TRPC5 Hs00202960_m1 TRPC6 Hs00175753_m1 TRPC7 Hs00220638_m1 TRPML1 Hs01100653_m1 TRPML2 Hs00401920_m1 TRPML3 Hs00217663_m1 TRPP1 Hs00165517_m1 TRPP2 Hs00175850_m1 TRPP3 Hs00950467_m1 Open in a separate window TaqManTM gene expression assays used in this study 2.4.

Techniques: Gene Expression

Journal: Frontiers in Pharmacology

Article Title: Hypo-Osmotic Loading Induces Expression of IL-6 in Nucleus Pulposus Cells of the Intervertebral Disc Independent of TRPV4 and TRPM7

doi: 10.3389/fphar.2020.00952

Figure Lengend Snippet: TaqMan primers list.

Article Snippet: TRPC4 , Transient Receptor Potential Cation Channel, Subfamily C, Member 4 , Bt03214662_m1.

Techniques: Activation Assay

Journal: Frontiers in Pharmacology

Article Title: Hypo-Osmotic Loading Induces Expression of IL-6 in Nucleus Pulposus Cells of the Intervertebral Disc Independent of TRPV4 and TRPM7

doi: 10.3389/fphar.2020.00952

Figure Lengend Snippet: Primary screening: number of donors (min. n = 0; max. n = 3), in which the mRNA expression of all available TRP channel was detectable using TaqMan gene array.

Article Snippet: TRPC4 , Transient Receptor Potential Cation Channel, Subfamily C, Member 4 , Bt03214662_m1.

Techniques: Expressing

Journal: Frontiers in Pharmacology

Article Title: Hypo-Osmotic Loading Induces Expression of IL-6 in Nucleus Pulposus Cells of the Intervertebral Disc Independent of TRPV4 and TRPM7

doi: 10.3389/fphar.2020.00952

Figure Lengend Snippet: Secondary screening: number of donors (min. n = 0; max. n = 5), in which the mRNA expression of all selected TRP channels was detectable using TaqMan gene array and qPCR.

Article Snippet: TRPC4 , Transient Receptor Potential Cation Channel, Subfamily C, Member 4 , Bt03214662_m1.

Techniques: Expressing