Journal: Applied and Environmental Microbiology

Article Title: Microbiological and Cellular Evaluation of a Fluorine-Phosphorus-Doped Titanium Alloy, a Novel Antibacterial and Osteostimulatory Biomaterial with Potential Applications in Orthopedic Surgery

doi: 10.1128/AEM.02271-18

Figure Lengend Snippet: Neutralization of the bactericidal effect of Al released from bNT Ti-6Al-4V on S. aureus 15981 and S. maltophilia ATCC 13637. (a) Percent viability of adhered bacteria. (b) Abundance of bacteria in solution after incubation. (c) Concentration of Al released from each alloy in the presence of planktonic bacteria (PB) and of Al not recruited by planktonic bacteria (filtered planktonic bacteria, FPB). *, P

Article Snippet: The bacterial viability of adhered bacteria of S. aureus 15981, S. epidermidis ATCC 35984, and S. maltophilia ATCC 13637 decreased significantly on bNT Ti-6Al-4V samples compared to levels on CP Ti-6Al-4V ( ) in a way that resembled the statistically significant decrease in the planktonic bacterial viability of S. maltophilia ATCC 13637 and the nonsignificant viability of S. aureus 15981 ( ).

Techniques: Neutralization, Incubation, Concentration Assay

Journal: International Journal of Medical Sciences

Article Title: Anti-biofilm and Anti-Virulence Efficacy of Celastrol Against Stenotrophomonas maltophilia

doi: 10.7150/ijms.23924

Figure Lengend Snippet: Effect of celastrol on motility of S. maltophilia . Motility assays were performed using 0.15% agar plate containing 1% tryptone and 0.5% NaCl. (A) ATCC 13637 and (B) GNU2233 strains were inoculated in the center of motility agar surface including DMSO 1% (v/v) or celastrol at sub-inhibitory concentrations. The inoculated plates were incubated at 37 °C and the diameter (mm) of circular swimming zone was measured at 24 h. Mean values of triplicate independent studies are shown. Error bars represent standard deviations (SDs). Asterisks (*) indicate statistically significant change (p

Article Snippet: Dispersion of established S. maltophilia biofilms by celastrol The biofilm dispersion activity of celastrol was investigated in S. maltophilia strains (ATCC 13637 and GNU2233) by measuring the biomass of biofilms.

Techniques: Incubation

Journal: International Journal of Medical Sciences

Article Title: Anti-biofilm and Anti-Virulence Efficacy of Celastrol Against Stenotrophomonas maltophilia

doi: 10.7150/ijms.23924

Figure Lengend Snippet: Transcriptional profiles of S. maltophilia cells treated with or without celastrol. ATCC 13637 and GNU 2233 were cultivated in CA-MHB medium at 37 °C for 6 h with shaking (250 rpm). Thereafter, they were further incubated at 37 °C for 4 h with or without celastrol at (A) 5 μg/mL (ATCC 13637) or (B) 20 μg/mL (GNU2233) without shaking. Transcriptional profiles were examined by qRT-PCR analysis. Relative gene expressions indicate transcriptional levels after treatment with celastrol versus DMSO treated control (value of 1.0). The tmRNA was used to normalize the expression levels of genes tested. All measurements were conducted with three individual cultures. Additionally, three replications were used for each qRT-PCR reaction. Mean values of triplicate independent studies are shown. Error bars represent standard deviations (SDs). Asterisks (*) indicate statistically significant change (p

Article Snippet: Dispersion of established S. maltophilia biofilms by celastrol The biofilm dispersion activity of celastrol was investigated in S. maltophilia strains (ATCC 13637 and GNU2233) by measuring the biomass of biofilms.

Techniques: Incubation, Quantitative RT-PCR, Expressing

Journal: International Journal of Medical Sciences

Article Title: Anti-biofilm and Anti-Virulence Efficacy of Celastrol Against Stenotrophomonas maltophilia

doi: 10.7150/ijms.23924

Figure Lengend Snippet: Effect of celastrol on established biofilms formed by S. maltophilia strains ((A) ATCC 13637 and (B) GNU2233). To determine the ability of celastrol to disrupt mature biofilms, S. maltophilia strains were cultured at 37 °C for 24 h to form established biofilms in microtitre plates. Celastrol was then added at different concentrations. These cultures were incubated further for 24 h. Total biofilm formation of S. maltophilia strains were examined by measuring absorbance at OD595nm. Mean values of triplicate independent studies are shown. Error bars represent standard deviations (SDs). Asterisks (*) indicate statistically significant change (p

Article Snippet: Dispersion of established S. maltophilia biofilms by celastrol The biofilm dispersion activity of celastrol was investigated in S. maltophilia strains (ATCC 13637 and GNU2233) by measuring the biomass of biofilms.

Techniques: Cell Culture, Incubation

Journal: International Journal of Medical Sciences

Article Title: Anti-biofilm and Anti-Virulence Efficacy of Celastrol Against Stenotrophomonas maltophilia

doi: 10.7150/ijms.23924

Figure Lengend Snippet: Effect of celastrol on metabolic activities of established biofilms formed by S. maltophilia strains ((A) ATCC 13637 and (B) GNU2233). The viability of cells within the biofilms treated with or without celastrol was evaluated by XTT viability assay. Specific absorbance indicates A490nm (Test) - A490nm (Blank) - A655nm (Test). Mean values of triplicate independent studies are shown. Error bars represent standard deviations (SDs). Asterisks (*) indicate statistically significant change (p

Article Snippet: Dispersion of established S. maltophilia biofilms by celastrol The biofilm dispersion activity of celastrol was investigated in S. maltophilia strains (ATCC 13637 and GNU2233) by measuring the biomass of biofilms.

Techniques: Viability Assay

Journal: International Journal of Medical Sciences

Article Title: Anti-biofilm and Anti-Virulence Efficacy of Celastrol Against Stenotrophomonas maltophilia

doi: 10.7150/ijms.23924

Figure Lengend Snippet: Microscopic evaluation of the effect of celastrol on established biofilms formed on glass surface using a confocal laser scanning microscope. CLSM micrographs of S. maltophilia ATCC 13637 biofilms treated with (A) DMSO 1 % (v/v) (control) and celastrol at concentration of (B) 40 μg/mL or (B) 80 μg/mL. Green (FITC) and red (Concanavalin A) colors indicate proteins and carbohydrates of EPS, respectively. Blue fluorescent staining (DAPI) represents extracellular DNA. CLSM images of panel (A) of DMSO-treated control S. maltophilia biofilms have spatial biomass distribution of mature biofilms and thick coating of biofilm with compact architecture characterized by large clumps. On the other hand, in panels (B) and (C) CLSM images of S. maltophilia biofilm treated with celastrol at 40 and 80 μg/mL, the established biofilms are almost disrupted and biofilm cells are remained as long cell chains. Magnification: 400x for each dose. Scale bar: 50 μm.

Article Snippet: Dispersion of established S. maltophilia biofilms by celastrol The biofilm dispersion activity of celastrol was investigated in S. maltophilia strains (ATCC 13637 and GNU2233) by measuring the biomass of biofilms.

Techniques: Laser-Scanning Microscopy, Confocal Laser Scanning Microscopy, Concentration Assay, Staining

Journal: International Journal of Medical Sciences

Article Title: Anti-biofilm and Anti-Virulence Efficacy of Celastrol Against Stenotrophomonas maltophilia

doi: 10.7150/ijms.23924

Figure Lengend Snippet: Effect of celastrol on protease production of S. maltophilia . Proteolytic activity was determined using casein agar. (A) ATCC 13637 and (B) GNU2233 cells were incubated with or without sub-inhibitory concentrations of celastrol at 37 °C for 24 h. Secreted protease assays were performed using culture supernatants. Diameters of transparent zones surrounding the holes were measured at 48 h. Mean values of triplicate independent studies are shown. Error bars represent standard deviations (SDs). Asterisks (*) indicate statistically significant change (p

Article Snippet: Dispersion of established S. maltophilia biofilms by celastrol The biofilm dispersion activity of celastrol was investigated in S. maltophilia strains (ATCC 13637 and GNU2233) by measuring the biomass of biofilms.

Techniques: Activity Assay, Incubation

Journal: International Journal of Medical Sciences

Article Title: Anti-biofilm and Anti-Virulence Efficacy of Celastrol Against Stenotrophomonas maltophilia

doi: 10.7150/ijms.23924

Figure Lengend Snippet: Effect of celastrol on biofilm formation of S. maltophilia . Biofilms produced by S. maltophilia strains ((A) ATCC 13637 and (B) GNU2233) were quantified by measuring the absorbance values at OD595nm in the presence or absence of celastrol at increasing doses. Planktonic cell growth of S. maltophilia strains ((C) ATCC 13637 and (D) GNU2233) in the presence of celastrol was determined by measuring absorbance at wavelength of 600 nm. Mean values of triplicate independent studies are shown. Error bars represent standard deviations (SDs). Asterisks (*) indicate statistically significant change (p

Article Snippet: Dispersion of established S. maltophilia biofilms by celastrol The biofilm dispersion activity of celastrol was investigated in S. maltophilia strains (ATCC 13637 and GNU2233) by measuring the biomass of biofilms.

Techniques: Produced

Journal: Molecules

Article Title: Ethanolic Extract of Folium Sennae Mediates the Glucose Uptake of L6 Cells by GLUT4 and Ca2+

doi: 10.3390/molecules23112934

Figure Lengend Snippet: The IP3R receptor is involved in FSE-stimulated intracellular Ca 2+ release. ( A ) Under extracellular 2 mM Ca 2+ , cells were treated with 100 μM 2-APB (IP3RS blocker) for 30 min and then treated with 60 μg/mL FSE. ( B ) Under extracellular 2 mM Ca 2+ , cells were incubated with 30 μM Ryanodine (RyR blocker) for 30 min, and the changes of IRAP-mOrange in the PM area and intracellular Ca 2+ were measured after stimulation with 60 μg/mL FSE. Significance analysis: * p

Article Snippet: Wortmannin, BAPTA-AM, U73122, Ryanodine, and 2-APB were purchased from Sigma (St. Louis, MO, USA).

Techniques: Incubation

Journal: Biochemistry

Article Title: Two Regions of the Ryanodine Receptor Calcium Channel Are Involved in Ca2+-Dependent Inactivation

doi: 10.1021/bi401586h

Figure Lengend Snippet: Two regions are involved in isoform-specificCa 2+ -dependentinactivation of RyRs. (A) Ca 2+ -dependent changes in theactivities of WT RyR1 (●) and WT RyR2 (○) were measuredin [ 3 H]ryanodine binding assays in the absence (left) orpresence

Article Snippet: [3 H]Ryanodine was obtained fromPerkinElmer (Waltham, MA) and unlabeled ryanodine from Calbiochem(La Jolla, CA).

Techniques: Binding Assay

Journal: Scientific Reports

Article Title: Dual action of L-Lactate on the activity of NR2B-containing NMDA receptors: from potentiation to neuroprotection

doi: 10.1038/s41598-018-31534-y

Figure Lengend Snippet: The RyRs blockade impacts only the maintenance of Ca 2+ signal during L-Lactate-induced potentiation. ( A 1 , B 1 ) Averaged Ca 2+ traces recorded from 65 neurons ( A 1 ) and 49 neurons ( B 1 ) following 2 successive applications of glutamate/glycine (2 min; dots) in control and in presence of L-Lactate (10 mM), the neurons in B 1 coming from a culture pre-treated with ryanodine (100 μM). ( A 2 , B 2 ) The scaling of evoked Ca 2+ responses obtained in ( A 1 and B 1 ) shows that the L-Lactate-induced potentiation also corresponds to an increase of the decay time (in comparison with the scaled Ca 2+ response obtained in control, A 2 ), with ryanodine blocking this effect on the Ca 2+ response kinetics ( B 2 ). ( C 1 , C 2 ) Bar charts summarizing the effect of L-Lactate on the Ca 2+ signal triggered by co-application of glutamate/glycine on untreated cultures (n cult = 11; n cells = 468; C 1 ) or cultures pre-treated with ryanodine (n cult = 7; n cells = 331; C 2 ). Note that, when cultures are pre-treated with ryanodine, the potentiating effect of L-Lactate significantly persists only for the amplitude of the Ca 2+ signal. Results are presented as means ± SEM (*p

Article Snippet: For experiments with Ryanodine (Tocris), the blocker is added to the loading solution at 100 μM during 30 minutes.

Techniques: Blocking Assay

Journal: Applied and Environmental Microbiology

Article Title: Application of Siderotyping for Characterization of Pseudomonas tolaasii and "Pseudomonas reactans" Isolates Associated with Brown Blotch Disease of Cultivated Mushrooms

doi:

Figure Lengend Snippet: Heterologous PVD-mediated 59 Fe incorporation by “ P. reactans ” strains belonging to the eight siderovars. Ordinate values correspond to 59 Fe radioactivity expressed in counts per minute as measured after 20 min of incubation (see Materials and Methods). Abscissa numbers 1 to 36 correspond to the structurally different PVDs tested, originating from the following bacterial strains (PVD numbers are given in parentheses): Pseudomonas strain E8 (1), P. syringae ATCC 19310 (2), P. fluorescens 9AW (3), P. putida ATCC 12633 (4), P. fluorescens 51W (5), P. aeruginosa Pa6 (6), P. fluorescens CCM 2798 (7), P. fluorescens CHA0 (8), P. tolaasii NCPPB 2192 (9), P. aeruginosa ATCC 27853 (10), P. fluorescens ii (11), P. fluorescens SB8.3 (12), P. fluorescens ATCC 17400 (13), P. fluorescens 1.3 (14), Pseudomonas strain 267 (15), P. fluorescens ATCC 13525 (16), P. aeruginosa ATCC 15692 (17), P. fluorescens strain 18.1 (18), P. fluorescens 12 (19), P. fluorescens CFBP 2392 (20), Pseudomonas strain L1 (21), Pseudomonas sp. strain ATCC 15915 (22), P. putida WCS358 (23), P. monteilii CFML 90-54 (24), “ P. mosselii ” CFML 90-77 (25), P. rhodesiae CFML 92-104 (26), P. veronii CFML 92-124 (27), Pseudomonas sp. strain CFML 90-33 (28), Pseudomonas sp. strain CFML 90-51 (29), Pseudomonas sp. strain CFML 90-52 (30), Pseudomonas sp. strain CFML 95-307 (31), Pseudomonas sp. strain 2908 (32), Pseudomonas sp. strain A214 (33), P. fluorescens PL7 (34), P. fluorescens PL8 (35), and the PVD synthesized by the strain under investigation (36). The counts per minute were corrected for the blank values obtained in assays without bacteria.

Article Snippet: 3 strains, which could be related to P. fluorescens ATCC 13525, the type strain of the P. fluorescens biovar I, but also to the P. chlororaphis species since P. chlororaphis ATCC 9446 produces the same PVD as P. fluorescens ATCC 13525 ( ).

Techniques: Radioactivity, Incubation, Synthesized

Journal: Applied and Environmental Microbiology

Article Title: Application of Siderotyping for Characterization of Pseudomonas tolaasii and "Pseudomonas reactans" Isolates Associated with Brown Blotch Disease of Cultivated Mushrooms

doi:

Figure Lengend Snippet: IEF profiles of PVDs for some “ P. reactans ” isolates and some other pseudomonads having similar uptake efficiencies. PVDs originated from the indicated strains: lane 1, “ P. reactans ” PL24.1 (sv. 1); lane 2, P. fluorescens ATCC 17400; lane 3, “ P. reactans ” PL10.9 (sv. 6); lane 4, P. fluorescens 51W; lane 5, P. monteilii CFML 90.54; lane 6, “ P. reactans ” PL4.14 (sv. 7); lane 7, P. fluorescens 12; lane 8, P. fluorescens 18.1; lane 9, P. aeruginosa ATCC 15692; lane 10, “ P. reactans ” PS11.4 (sv. 2); lane 11, “ P. reactans ” PS3b (sv. 5); lane 12, P. rhodesiae CFML 92.104; lane 13, P. veronii CFML 92.124; lane 14, “ P. reactans ” PS7.2 (sv. 3); lane 15, P. fluorescens ATCC 13525.

Article Snippet: 3 strains, which could be related to P. fluorescens ATCC 13525, the type strain of the P. fluorescens biovar I, but also to the P. chlororaphis species since P. chlororaphis ATCC 9446 produces the same PVD as P. fluorescens ATCC 13525 ( ).

Techniques: Electrofocusing

Journal: Brain and Behavior

Article Title: Ryanodine‐ and Ca MKII‐dependent release of endogenous CGRP induces an increase in acetylcholine quantal size in neuromuscular junctions of mice. Ryanodine‐ and CaMKII‐dependent release of endogenous CGRP induces an increase in acetylcholine quantal size in neuromuscular junctions of mice

doi: 10.1002/brb3.1058

Figure Lengend Snippet: An increase in MEPP amplitude in the presence of ryanodine. (a) Representative intracellular recordings of MEPP s in control and in the presence of ryanodine (0.1 μM). Insets show the enlarged views of MEPP s. (b) Mean amplitude of MEPP s in control ( n = 23), during first 30 min of ryanodine application ( n = 15) and within 30–90 min of ryanodine application ( n = 28). * p

Article Snippet: Ryanodine (an activator of RyR) and an inhibitor of protein kinase A H‐89 were obtained from Enzo Life Sciences (Farmingdale, NY, USA).

Techniques:

Journal: Brain and Behavior

Article Title: Ryanodine‐ and Ca MKII‐dependent release of endogenous CGRP induces an increase in acetylcholine quantal size in neuromuscular junctions of mice. Ryanodine‐ and CaMKII‐dependent release of endogenous CGRP induces an increase in acetylcholine quantal size in neuromuscular junctions of mice

doi: 10.1002/brb3.1058

Figure Lengend Snippet: CGRP mediates the ryanodine‐induced increase in MEPP amplitude. (a) Mean amplitude of MEPP s in control ( n = 16) and during application of exogenous CGRP (10 nM ) for 40 min ( n = 18). * p

Article Snippet: Ryanodine (an activator of RyR) and an inhibitor of protein kinase A H‐89 were obtained from Enzo Life Sciences (Farmingdale, NY, USA).

Techniques:

Journal: Brain and Behavior

Article Title: Ryanodine‐ and Ca MKII‐dependent release of endogenous CGRP induces an increase in acetylcholine quantal size in neuromuscular junctions of mice. Ryanodine‐ and CaMKII‐dependent release of endogenous CGRP induces an increase in acetylcholine quantal size in neuromuscular junctions of mice

doi: 10.1002/brb3.1058

Figure Lengend Snippet: The potentiating effect of ryanodine on the amplitude of MEPP s is mediated by Ca MKII . (a) Mean amplitude of MEPP s in control ( n = 15 when only KN ‐62 was applied subsequently and n = 16 when ryanodine was applied in the presence of KN ‐62), during application of 3 μM KN ‐62 ( n = 18) and ryanodine (0.1 μM) in the presence of KN ‐62 ( n = 26). Histograms and error bars indicate the mean and SEM , respectively. (b) Cumulative probability plots of MEPP amplitudes in control and in the presence of KN ‐62, p = 0.42. (c) Cumulative probability plots of MEPP amplitudes in control and during application of ryanodine in the presence of KN ‐62, p = 0.11. (d) Mean amplitude of MEPP s in control ( n = 16 when only KN ‐93 was applied subsequently and n = 17 when ryanodine was applied in the presence of KN ‐93), during application of 3 μM KN ‐93 ( n = 21) and ryanodine (0.1 μM) in the presence of KN ‐93 ( n = 31). Histograms and error bars indicate the mean and SEM , respectively. (e) Cumulative probability plots of MEPP amplitudes in control and in the presence of KN ‐93, p = 0.12. (f) Cumulative probability plots of MEPP amplitudes in control and during application of ryanodine in the presence of KN ‐93, p = 0.14. (g) Mean amplitude of MEPP s in control ( n = 19 when only KN ‐92 was applied subsequently and n = 21 when ryanodine was applied in the presence of KN ‐92), during application of 3 μM KN ‐92 ( n = 26) and ryanodine (0.1 μM) in the presence of KN ‐92 ( n = 26). Histograms and error bars indicate the mean and SEM , respectively. (e) Cumulative probability plots of MEPP amplitudes in control and in the presence of KN ‐92, p = 0.58. (f) Cumulative probability plots of MEPP amplitudes in control and during application of ryanodine in the presence of KN ‐92, p

Article Snippet: Ryanodine (an activator of RyR) and an inhibitor of protein kinase A H‐89 were obtained from Enzo Life Sciences (Farmingdale, NY, USA).

Techniques:

Journal: Brain and Behavior

Article Title: Ryanodine‐ and Ca MKII‐dependent release of endogenous CGRP induces an increase in acetylcholine quantal size in neuromuscular junctions of mice. Ryanodine‐ and CaMKII‐dependent release of endogenous CGRP induces an increase in acetylcholine quantal size in neuromuscular junctions of mice

doi: 10.1002/brb3.1058

Figure Lengend Snippet: Intracellular mechanisms of ryanodine action at the mouse NMJ s. (a) Vesicular AC h transporter blocker vesamicol (1 μM) prevented MEPP amplitude increase induced by ryanodine (0.1 μM). Histograms and error bars indicate the mean and SEM , respectively. (b) The overlapping of cumulative probability plots of MEPP amplitudes in control and during application of ryanodine in the presence of vesamicol, p = 0.78. (c) Inhibition of PKA by H‐89 (0.1 μM) prevents the ryanodine‐induced increase in MEPP amplitude. Histograms and error bars indicate the mean and SEM , respectively. (d) Cumulative probability plots of MEPP amplitudes in control and during application of ryanodine in the presence of H‐89, p = 0.60

Article Snippet: Ryanodine (an activator of RyR) and an inhibitor of protein kinase A H‐89 were obtained from Enzo Life Sciences (Farmingdale, NY, USA).

Techniques: Inhibition

Journal: Brain and Behavior

Article Title: Ryanodine‐ and Ca MKII‐dependent release of endogenous CGRP induces an increase in acetylcholine quantal size in neuromuscular junctions of mice. Ryanodine‐ and CaMKII‐dependent release of endogenous CGRP induces an increase in acetylcholine quantal size in neuromuscular junctions of mice

doi: 10.1002/brb3.1058

Figure Lengend Snippet: Ryanodine effect on single evoked EPP s and MEPP s. (a) Representative recordings of EPP s (left) and MEPP s (right) in control and in the presence of ryanodine for 30–90 min. (b) Mean EPP s amplitude, MEPP s amplitude, and quantal content of EPP s (left to right) in control ( n = 40), during first 30 min of ryanodine application ( n = 18) and within 30–90 min of ryanodine application ( n = 27). * p

Article Snippet: Ryanodine (an activator of RyR) and an inhibitor of protein kinase A H‐89 were obtained from Enzo Life Sciences (Farmingdale, NY, USA).

Techniques:

Journal: Journal of Applied Physiology

Article Title: Chronic low-intensity exercise attenuates cardiomyocyte contractile dysfunction and impaired adrenergic responsiveness in aortic-banded mini-swine

doi: 10.1152/japplphysiol.00840.2017

Figure Lengend Snippet: Protein expression of Ca 2+ handling proteins. A : example Western blots of sarco/endoplasmic reticulum Ca 2+ ATPase (SERCA; 100 kDa), phospholamban (PLB; 6 kDa), the Ser16 phosphorylation site of PLB (SER16-PLB; 6 kDa), the Thr17 phosphorylation site of PLB (THR17-PLB; 6 kDa), the Na + -K + -ATPase (NA/K; 100 kDa), sodium-calcium exchanger (NCX; 120 kDa), ryanodine receptor (RYR; 565 kDa), and calsequestrin (CSQ; 50 kDa). Summary data of protein expression (relative to CSQ) of SERCA ( B ), PLB ( C ), SERCA/PLB ratio ( D ), SER16-PLB ( E ), THR17-PLB ( F ), Na + -K + ATPase ( G ), NCX ( H ), and RYR ( I ). Data shown are from whole heart homogenates of CON, AB, and AB-LIT groups; n = 5–7 animals per group. * P

Article Snippet: Blots were incubated overnight (25°C) with the primary antibody against SERCA2a (1:1,000; ThermoFisher Scientific), phospholamban (PLB; 1:1,000; ThermoFisher Scientific), sodium-calcium exchanger (NCX; 1:1,000; ThermoFisher Scientific), Na+ -K+ -ATPase (1:1,000; ThermoFisher Scientific), ryanodine receptor (RyR; 1:1,000; ThermoFisher Scientific), Ser16 phosphorylated PLB (1:1,000; Badrilla), Thr17 phosphorylated PLB (1:1,000; Badrilla), and calsequestrin (CSQ; 1:10,000; ThermoFisher Scientific).

Techniques: Expressing, Western Blot

Journal: The Journal of Neuroscience

Article Title: Ca2+-Dependent Regulation of Rho GTPases Triggers Turning of Nerve Growth Cones

doi: 10.1523/JNEUROSCI.4889-04.2005

Figure Lengend Snippet: PKC mediates Ca 2+ signaling to Rho GTPases. a , b , Western blots showing the active form of PKC and CaMKII in HEK293T cells or cultured cerebellar granule cells treated with ryanodine (10 n m ) at different times. Western blots using antibody against actin

Article Snippet: Ryanodine was from Alomone Laboratories (Jerusalem, Israel), and fluo-4 AM and fura-Red AM were from Molecular Probes (Eugene, OR).

Techniques: Western Blot, Cell Culture

Journal: The Journal of Neuroscience

Article Title: Ca2+-Dependent Regulation of Rho GTPases Triggers Turning of Nerve Growth Cones

doi: 10.1523/JNEUROSCI.4889-04.2005

Figure Lengend Snippet: Ryanodine-induced attractive turning response depends on both Ca 2+ and Rho GTPases. a , Distribution of growth cone turning angles (left) for control neurons and neurons treated with BAPTA-AM (BAPTA; 10 μ m ), thapsigargin (Thaps; 10 μ m ),

Article Snippet: Ryanodine was from Alomone Laboratories (Jerusalem, Israel), and fluo-4 AM and fura-Red AM were from Molecular Probes (Eugene, OR).

Techniques:

Journal: The Journal of Neuroscience

Article Title: Ca2+-Dependent Regulation of Rho GTPases Triggers Turning of Nerve Growth Cones

doi: 10.1523/JNEUROSCI.4889-04.2005

Figure Lengend Snippet: Ca 2+ elevation regulates the activity of Rho GTPases in transfected HEK293T cells. a , b , Representative Western blots showing the dose-response and time course of the activation of Cdc42 and Rac and inactivation of RhoA by bath-applied ryanodine in HEK293T

Article Snippet: Ryanodine was from Alomone Laboratories (Jerusalem, Israel), and fluo-4 AM and fura-Red AM were from Molecular Probes (Eugene, OR).

Techniques: Activity Assay, Transfection, Western Blot, Activation Assay

Journal: The Journal of Neuroscience

Article Title: Ca2+-Dependent Regulation of Rho GTPases Triggers Turning of Nerve Growth Cones

doi: 10.1523/JNEUROSCI.4889-04.2005

Figure Lengend Snippet: Ryanodine-induced attraction depends on CaMKII and PKC. Distribution of turning angles in a gradient of ryanodine ( a , c , 10 μ m in the pipette) or netrin-1 ( b , 5 μg/ml in the pipette) for cultured Xenopus spinal neurons in control condition

Article Snippet: Ryanodine was from Alomone Laboratories (Jerusalem, Israel), and fluo-4 AM and fura-Red AM were from Molecular Probes (Eugene, OR).

Techniques: Transferring, Cell Culture

Journal: The Journal of Neuroscience

Article Title: Ca2+-Dependent Regulation of Rho GTPases Triggers Turning of Nerve Growth Cones

doi: 10.1523/JNEUROSCI.4889-04.2005

Figure Lengend Snippet: Ca 2+ -dependent regulation of Rho GTPase activity in cultured neurons. a , Activation of Cdc42 and inactivation of RhoA by bath-applied ryanodine (10 n m . The effects

Article Snippet: Ryanodine was from Alomone Laboratories (Jerusalem, Israel), and fluo-4 AM and fura-Red AM were from Molecular Probes (Eugene, OR).

Techniques: Activity Assay, Cell Culture, Activation Assay

Journal: The Journal of Neuroscience

Article Title: Ca2+-Dependent Regulation of Rho GTPases Triggers Turning of Nerve Growth Cones

doi: 10.1523/JNEUROSCI.4889-04.2005

Figure Lengend Snippet: Growth cone turning and local [Ca 2+ ] i elevation induced by ryanodine gradient. a , Microscopic images of a cultured Xenopus spinal neuron at the beginning (0 min) and the end (60 min) at a 1 hexposure to a ryanodine gradient created by pulsatile application

Article Snippet: Ryanodine was from Alomone Laboratories (Jerusalem, Israel), and fluo-4 AM and fura-Red AM were from Molecular Probes (Eugene, OR).

Techniques: Cell Culture

Journal: The Journal of Neuroscience

Article Title: Ca2+-Dependent Regulation of Rho GTPases Triggers Turning of Nerve Growth Cones

doi: 10.1523/JNEUROSCI.4889-04.2005

Figure Lengend Snippet: Regulation of the activities of Rho GTPases by ryanodine requires CaMKII and PKC. a , b , Regulation of Rho GTPases by bath-applied ryanodine (10 n m for 3 min) in HEK293T cells transfected with wild-type Rho GTPases was abolished by specific inhibitors

Article Snippet: Ryanodine was from Alomone Laboratories (Jerusalem, Israel), and fluo-4 AM and fura-Red AM were from Molecular Probes (Eugene, OR).

Techniques: Transfection

Journal: The Journal of Physiology

Article Title: Size-dependent heterogeneity of contractile Ca2+ sensitization in rat arterial smooth muscle

doi: 10.1113/jphysiol.2012.241315

Figure Lengend Snippet: Effect of treatment with 1 μ m ryanodine (rye, A ) and 1 μ m nicardipine (nic, B ) on time course of Ca 2+ rise in response to 30 μ m PE in small mesenteric artery

Article Snippet: To deplete SR Ca2+ stores, arterial rings were incubated in normal external solution containing 1 μ m ryanodine (BioMol, Plymouth Meeting, PA, USA) and 20 m m caffeine for 15 min and washed with the same solution without caffeine for another 15 min whereupon caffeine no longer evoked a transient contraction ( ).

Techniques:

Journal: The Journal of Physiology

Article Title: Size-dependent heterogeneity of contractile Ca2+ sensitization in rat arterial smooth muscle

doi: 10.1113/jphysiol.2012.241315

Figure Lengend Snippet: Effect of 1 μ m ryanodine, 1 μ m nicardipine, and a combination of the two blockers on 30 μ m PE-induced contraction in mesenteric ( A ; n = 3–6), caudal ( B ; n = 4–6) and aortic arteries ( C ; n = 4)

Article Snippet: To deplete SR Ca2+ stores, arterial rings were incubated in normal external solution containing 1 μ m ryanodine (BioMol, Plymouth Meeting, PA, USA) and 20 m m caffeine for 15 min and washed with the same solution without caffeine for another 15 min whereupon caffeine no longer evoked a transient contraction ( ).

Techniques:

Journal: Developmental cell

Article Title: Intracellular calcium mobilization is required for Sonic hedgehog signaling

doi: 10.1016/j.devcel.2018.04.013

Figure Lengend Snippet: RyR function is required for Shh-dependent Gli-mediated gene expression. (A–H) Dorsal view images of live Tg(8xGli:mCherry-NLS-Odc1) embryos at 12hpf (A,C,E,G) or 24hpf (B,D,F,H) that had been treated with vehicle (0.5%DMSO), cyclopamine, azumolene, or ryanodine. At 12hpf, the position of the notochord (ntc) just ventral to the FP is outlined by dashed lines. (A) In control 12hpf embryos, mCherry is expressed in nuclei of cells responding to Shh, including adaxial cells (arrowhead) and FP cells (arrow). (B) At 24hpf, mCherry is expressed in nuclei of slow muscle cells (arrowhead) and cells in the ventral neural tube (arrow). (C–H) mCherry expression is reduced in embryos treated with each drug. (I) Tail-transected Tg(8xGli:mCherry-NLS-Odc1) embryos were soaked in vehicle or 4-CmC from 16 to 18hpf, fixed, and imaged. (J–L) Potentiation of RyR channel activity with 4-CmC treatment results in increased numbers of presumptive slow muscle nuclei that express the mCherry reporter (J and K are lateral views, arrowhead indicates nuclei of slow muscle cells and arrow indicates cells in the ventral neural tube). (L) Quantification of mCherry + nuclei per somite in 4-CmC-treated embryos. Each point represents a single somite and the horizontal line represents the mean. (M–Q) RyR activity affects endogenous ptch2 expression as detected by whole mount in situ hybridization in 24hpf embryos. (M–Q) Lateral views reveal expression in somites and (M′–Q′) transverse sections reveal expression in slow muscle cells surrounding the notochord and in the ventral neural tube. As compared with WT embryos, ptch2 expression is diminished azumolene-treated and MZryr1a (−/−) ;MZryr2a (−/−) ;MZryr3 (−/−) mutant embryos, and it is enhanced in 4-CmC-treated embryos. Scale bars indicate 25 μm.

Article Snippet: Ryanodine (Santa Cruz Biotechnology), azumolene (Santa Cruz Biotechnology), NAC (Sigma-Aldrich), and thapsigargin (Sigma-Aldrich) were prepared in DMSO; aldrithiol (DTDP, Sigma-Aldrich) and cyclopamine (LC Labs) were prepared in ethanol; and tricaine (MS-222, Sigma) and 4-CmC were prepared in water.

Techniques: Expressing, Activity Assay, In Situ Hybridization, Mutagenesis

Journal: Developmental cell

Article Title: Intracellular calcium mobilization is required for Sonic hedgehog signaling

doi: 10.1016/j.devcel.2018.04.013

Figure Lengend Snippet: RyR function is required for development of Shh-dependent neural crest-derived neurons of the dorsal root ganglia (DRGs) and the enteric nervous system (ENS). (A–L) Lateral views of live 72hpf Tg(isl2b:GFP) embryos with GFP-labeled Rohon Beard neurons (arrow) and DRGs (arrowhead). Embryos were treated between 24 and 48hpf with cyclopamine (cyc), azumolene (azum), ryanodine (ryan), N-acetyl cysteine (NAC), aldrithiol, thapsigargin (thaps), or tricaine at indicated concentrations or injected at the one-cell stage with ryr1a and ryr3 MOs. Arrowheads in C and G indicate the presence of small, faint DRGs. (M) Quantification of embryos treated as in A–L. DRGs present in somites 11–15 were counted. For each condition, the number of embryos analyzed is indicated. Comparisons are to control vehicle-treated embryos unless otherwise indicated. Data are represented as mean ±SEM. (N–Q) Lateral views of live 78hpf Tg(phox2bb:GFP) embryos with the ENS (arrow) labeled by GFP in control, cyclopamine-treated, ryr1a,3 .

Article Snippet: Ryanodine (Santa Cruz Biotechnology), azumolene (Santa Cruz Biotechnology), NAC (Sigma-Aldrich), and thapsigargin (Sigma-Aldrich) were prepared in DMSO; aldrithiol (DTDP, Sigma-Aldrich) and cyclopamine (LC Labs) were prepared in ethanol; and tricaine (MS-222, Sigma) and 4-CmC were prepared in water.

Techniques: Derivative Assay, Labeling, Injection