Journal: PLoS ONE

Article Title: A Host Defense Mechanism Involving CFTR-Mediated Bicarbonate Secretion in Bacterial Prostatitis

doi: 10.1371/journal.pone.0015255

Figure Lengend Snippet: (A)When 1×10 4 CFU of E.coli was inoculated to the apical compartment of the rat prostate epithelial cells for 18 h, there was no bacterial activity detected in the culture medium. 10 µM CFTR inh -172 (A), 1∶500 CFTR antibody (B) or 50 µM acetazolamide (C) were added with 1×10 5 E.coli to block CFTR or CAII activity and their effect on bacterial activity 18 hours after incubation was shown. (**P<0.01, ***P<0.001).

Article Snippet: For blocking of CFTR or carbonic anhydrase, the E coli treated epithelial cells were preincubated with 10 µM CFTR inh -172 (Sigma), a specific CFTR channel blocker or anti-CFTR antibody (1∶500, Alomone labs) or 50 µM Acetazolamide (Sigma), a carbonic anhydrase inhibitor for 24 h before the addition of bacteria.

Techniques: Activity Assay, Blocking Assay, Incubation

Journal: PLoS ONE

Article Title: A Host Defense Mechanism Involving CFTR-Mediated Bicarbonate Secretion in Bacterial Prostatitis

doi: 10.1371/journal.pone.0015255

Figure Lengend Snippet: (A) Comparison of E coli bacterial activities recovered from rat prostatitis models without or with CFTR inh -172 (10 µM). Each point indicates the bacterial CFU per gram of prostate tissue weight (***P<0.001). (B) E.coli up-regulated the expression of cytokine genes, CFTR and CAII in rat prostate as determined by RT-PCR. Data were from three experiments. (C) Expression of CFTR (160 kD) and CAII (29 kD) protein was significantly up-regulated in E.coli -infected rat prostate as determined by western blot. Data were from three experiments. (*P<0.05, **P<0.01, ***P<0.001).

Article Snippet: For blocking of CFTR or carbonic anhydrase, the E coli treated epithelial cells were preincubated with 10 µM CFTR inh -172 (Sigma), a specific CFTR channel blocker or anti-CFTR antibody (1∶500, Alomone labs) or 50 µM Acetazolamide (Sigma), a carbonic anhydrase inhibitor for 24 h before the addition of bacteria.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Infection, Western Blot

Journal: Frontiers in Physiology

Article Title: A Calcium-Dependent Chloride Current Increases Repetitive Firing in Mouse Sympathetic Neurons

doi: 10.3389/fphys.2018.00508

Figure Lengend Snippet: The chloride channel blocker 9AC diminishes firing frequency in both Ph and T neurons. (A) Repetitive firing of a Ph (left) neuron in control solution after the injection of 0.5 nA depolarizing current (black upper traces) and its modification after the addition of 2 mM 9AC (blue lower traces). The same effect in a T neuron (right). The injected current protocol is shown in each recording below the membrane potential. The RMP of cells is indicated at the beginning of each recording. Calibration bars apply to both panels. (B) Significant reduction of firing frequency elicited with 1.0 nA depolarizations in the presence of 2 mM 9AC for both populations of Ph ( n = 9) and T sympathetic neurons ( n = 9). Paired t -test: ∗ P < 0.05; ∗∗ P < 0.01.

Article Snippet: The Cl - channel blocker 9AC (Sigma, United States) was diluted in NaOH (1 M, and adjusted to pH 7.4 with HCl) and added to the control solution at a final concentration of 2 mM.

Techniques: Injection, Modification

Journal: Frontiers in Physiology

Article Title: A Calcium-Dependent Chloride Current Increases Repetitive Firing in Mouse Sympathetic Neurons

doi: 10.3389/fphys.2018.00508

Figure Lengend Snippet: The chloride channel blocker 9AC reduces firing frequency and instantaneous frequency in almost all sympathetic neurons. (A) The reduction in firing frequency was observed for all the amplitudes of the depolarizing pulse used (black circles for control solution and blue circles for 2 mM 9AC solution). Only cells firing two or more spikes in the control were considered (numbers in brackets). (B) The number of APs elicited is larger in control than in 9AC in the majority of cases (15 out of 18) with 0.5 nA depolarizing pulses. (C) The instantaneous frequency of spikes was reduced for all of the five first intervals between consecutive spikes studied with depolarizations of 1.0 nA. Symbols are the same as in A . The number of cells studied in each case is shown in brackets. (D) Effects of CaCC blockade with 9AC on firing frequency separately in cells with (types A and B) and without (type C) ADP for 1.0 nA depolarizing pulses. Paired t -test: ∗∗ P < 0.01; ∗∗∗ P < 0.001 (A,C) and Student’s t -test: ∗ P < 0.05 (D) .

Article Snippet: The Cl - channel blocker 9AC (Sigma, United States) was diluted in NaOH (1 M, and adjusted to pH 7.4 with HCl) and added to the control solution at a final concentration of 2 mM.

Techniques:

Journal: Frontiers in Physiology

Article Title: A Calcium-Dependent Chloride Current Increases Repetitive Firing in Mouse Sympathetic Neurons

doi: 10.3389/fphys.2018.00508

Figure Lengend Snippet: Anthracene-9′-carboxylic acid (9AC) abolishes ADPs without affecting synaptic transmission. (A) Representative recordings of ADPs were evoked after a train of 35 APs in control solution (black traces). ADPs disappeared completely (top panel) or were significantly reduced (low panel) after the addition of 2 mM 9AC (blue traces). Recordings of top panels are from the same cell before and after application of 9AC. The same applies to low panels. APs were evoked with short (5 ms) intracellular current pulses at 35 Hz shown in each recording below the membrane potential. Spikes are truncated at the top. Calibration bars apply to all panels. (B) Representative recording of an excitatory post-synaptic potential (EPSP) elicited with preganglionic stimulus in the refractory period of the AP of one cell in control solution. Addition of the chloride channel blocker 2 mM 9AC does not significantly modify the amplitude of EPSP. Two recordings are overimposed in B , one without stimulation of the preganglionic trunk (black and blue traces for control and 9AC, respectively) and the other with its stimulation and the evoked EPSP (red traces). Calibration bars apply to both panels. APs were evoked with a short (5 ms) intracellular current pulse shown in each recording below the membrane potential. The RMP of the cells is indicated at the beginning of each recording.

Article Snippet: The Cl - channel blocker 9AC (Sigma, United States) was diluted in NaOH (1 M, and adjusted to pH 7.4 with HCl) and added to the control solution at a final concentration of 2 mM.

Techniques: Transmission Assay

Journal: Frontiers in Physiology

Article Title: A Calcium-Dependent Chloride Current Increases Repetitive Firing in Mouse Sympathetic Neurons

doi: 10.3389/fphys.2018.00508

Figure Lengend Snippet: Anthracene-9′-carboxylic acid (9AC) affects only ADP.

Article Snippet: The Cl - channel blocker 9AC (Sigma, United States) was diluted in NaOH (1 M, and adjusted to pH 7.4 with HCl) and added to the control solution at a final concentration of 2 mM.

Techniques: