Journal: Cell reports

Article Title: Optogenetic Activation of Striatopallidal Neurons Reveals Altered HCN Gating in DYT1 Dystonia.

doi: 10.1016/j.celrep.2020.107644

Figure Lengend Snippet: Figure 7. ConcanamycinA Treatment Modulates HCN1 and HCN2 Expression (A) Cartoon showing the mechanism of concanamycinA (concan) treatment upon vesicular release. (B) sIPSCs recorded in slices of Tor1a+/+ and Tor1a+/DGAG treated with saline (black traces) or with concan (red traces). (C) Boxplot summarizes the effect of concan on firing frequency in Tor1a+/+ and Tor1a+/DGAG. (D) Western blots showing immunoreactivity of HCN1 (untreated N = 5, treated N = 7) and HCN2 (untreated N = 6, treated N = 6) subunits from GPe extracts from slices of animals treated with saline or concan. Protein values were normalized to the corresponding expression of tubulin and actin, respectively. Bar plots illustrate the effect of treatment on HCN subunits level in Tor1a+/+ and Tor1a+/DGAG mice. Levels were normalized to the control (CTRL) level of Tor1a+/+. (E) Cell-attached recordings of autonomous pacemaking activity of GP neurons from Tor1a+/DGAG treated with control solution (black) or concan (red). Boxplots show effect of concan on firing frequency and CV. (F) Whole-cell recordings from GP neurons of Tor1a+/DGAG during optical striatal stimulation (blue lines, dual pulse, 20 Hz) after treatment with concan. Five superimposed traces were aligned on the striatal stimulation. Peri-stimulus time histogram (PSTH) shows firing pause and resetting phase triggered by striatal stimulation. Each dot represents single measurements.

Article Snippet: Protein extracts were stored at 80 C and 10 mg of each sample, in Page LDS sample buffer 4x (Invitrogen) was analyzed to detect: TA (1:1000, Abcam AB_2240792), PV (1:2000, Abcam AB_956206), GAD67 (1:2000, Sigma Aldrich AB_261978), GAD65 (1:1000, Sigma Aldrich AB_10762670), VGAT (1:1000 SySy AB_887871), Gephyrin (1:500, Proteintech AB_2247770), HCN2 (1:1000, Proteintech AB_10951108) HCN1 (1:1000, Proteintech AB_11182929), rabbit anti-D2R (1:1000, Millipore AB_2094980), antiactin (Sigma Aldrich AB_476744) as loading control.

Techniques: Expressing, Saline, Western Blot, Control, Activity Assay

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: The Effects of Acupuncture on Bladder Interstitial Cells of Cajal Excitability in Rats with Overactive Bladder

doi: 10.1155/2013/261217

Figure Lengend Snippet: Relative expression level of the HCN2 mRNA in bladder determined by RT-PCR. ∗ versus normal control group P < 0.05. △ versus model group P < 0.05.

Article Snippet: Themembranes were blocked with 5% nonfat milk overnight at 4°C, and thenincubated with the primary antibodies recognizing β -actin (mouse monoclonal, 1 : 5000, Abcam, Cambridge, UK) or HCN2 antibody (rabbit polyclonal,1 : 1000, Cell Signaling Technology, Danvers, MA, USA) for 2 h at 22°C.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Control

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: The Effects of Acupuncture on Bladder Interstitial Cells of Cajal Excitability in Rats with Overactive Bladder

doi: 10.1155/2013/261217

Figure Lengend Snippet: Relative expression level of the HCN2 protein in bladder determined by Western blotting. ∗ versus normal control group P < 0.05. △ versus model group P < 0.05.

Article Snippet: Themembranes were blocked with 5% nonfat milk overnight at 4°C, and thenincubated with the primary antibodies recognizing β -actin (mouse monoclonal, 1 : 5000, Abcam, Cambridge, UK) or HCN2 antibody (rabbit polyclonal,1 : 1000, Cell Signaling Technology, Danvers, MA, USA) for 2 h at 22°C.

Techniques: Expressing, Western Blot, Control

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: The Effects of Acupuncture on Bladder Interstitial Cells of Cajal Excitability in Rats with Overactive Bladder

doi: 10.1155/2013/261217

Figure Lengend Snippet: The quantity of HCN2 channel in bladder ICCs. ((a), (b), (c), and (d)) Double-labeled immunofluorescence of HCN2 and c-Kit in bladder. Nucleus stained by DAPI (blue), c-Kit (green), and HCN2 (red) staining in urothelium, suburothelium, and muscle layer. Some ICCs were c-kit positive (↑), some were HCN2 positive (◆), and some were double labeled (▲). (a) Normal control group, (b) model group, (c) acupuncture group, (d) Glivec group. Scale bar is 50 μ m. (e) Comparisons of relative quantity of HCN2 channel in bladder ICCs after treatment. ∗ versus normal control group P < 0.05. △ versus model group P < 0.05.

Article Snippet: Themembranes were blocked with 5% nonfat milk overnight at 4°C, and thenincubated with the primary antibodies recognizing β -actin (mouse monoclonal, 1 : 5000, Abcam, Cambridge, UK) or HCN2 antibody (rabbit polyclonal,1 : 1000, Cell Signaling Technology, Danvers, MA, USA) for 2 h at 22°C.

Techniques: Labeling, Immunofluorescence, Staining, Control