p2x7r  (Alomone Labs)

Journal: Toxins

Article Title: Gambierol Blocks a K+ Current Fraction without Affecting Catecholamine Release in Rat Fetal Adrenomedullary Cultured Chromaffin Cells

doi: 10.3390/toxins14040254

Figure Lengend Snippet: Time-course, kinetics and concentration-dependent action of gambierol on outward K + current in rat fetal AMC cells. ( A ) Time course of 20 nM gambierol action on the K + current measured at the end of 90 ms depolarizing pulses to +40 mV from a holding potential of −70 mV, applied every 10 s (see schema in ( B )). The red arrow indicates the time of polyether addition to the bath. ( B ) Superimposed K + current recorded every 10-s pulsing, before and after the perfusion of gambierol (20 nM), during 90 ms depolarizing steps to +40 mV from a holding potential of −70 mV (schema). ( C ) Averaged normalized K + current recorded during 90 ms depolarizing steps to +40 mV from a holding potential of −70 mV (schema), under control conditions (black tracing), and after 20 nM gambierol (red tracing), note the slowing of the K + current activation. Data obtained from the same cell. The arrows indicate the activation time constants. ( D ) Concentration-response curve for the effect of gambierol on the steady-state K + current, measured after 90 ms depolarization steps to +40 mV from −70 mV. Each value, determined in the presence of 0.1–100 nM gambierol and normalized to its control value, represents the mean ± SEM of data obtained from 3–4 experiments. The theoretical curve was calculated as described in the text, with I SS , IC 50 and n H values of 42%, 5.8 nM and 0.89 (r 2 = 0.982), respectively. The external medium in A–D contained 1 µM TTX, 200 µM glibenclamide and 1 mM Cd 2+ to block, respectively, the Na V , K ATP and Ca V channels and prevent, indirectly, the activation of K Ca channels.

Article Snippet: Tetrodotoxin, apamin, iberiotoxin, and glibenclamide were purchased from Alomone Labs (Jerusalem, Israel), and Latoxan (Portes-lès-Valence, France).

Techniques: Concentration Assay, Activation Assay, Blocking Assay

Journal: Toxins

Article Title: Gambierol Blocks a K+ Current Fraction without Affecting Catecholamine Release in Rat Fetal Adrenomedullary Cultured Chromaffin Cells

doi: 10.3390/toxins14040254

Figure Lengend Snippet: The contribution of various current components (K Ca , K ATP ) and the action of gambierol (100 nM) on total K + current of rat fetal AMC cells. The K + current was recorded using the perforated whole-cell voltage-clamp technique, during 90 ms depolarizing steps from a holding potential of −70 mV, under the conditions indicated in the figure ( A , B ). Note in ( A , B ) the different sequences of gambierol addition to the external medium. ( a ) Superimposed traces of outward K + currents recorded under control conditions and after the perfusion of the different pharmacological agents indicated. AGI denotes the simultaneous perfusion of 400 nM apamin, 200 µM glibenclamide and 100 nM iberiotoxin. ( b ) Current-voltage relationships. The current was measured at the end of the depolarizing steps and expressed as a percentage of control values for the indicated agents. ( c ) Relative outward K + current contribution (expressed as a percentage of the total current) for the pharmacological treatments indicated. The K + current was measured at the end of the depolarizing steps to +40 mV and expressed as a percentage of control values. Note that the percentage of blocked K Ca and K ATP current components were not significantly different whatever the order of gambierol-induced channel inhibition was. In ( b , c ), data represent the mean ± SEM of 4 different experiments.

Article Snippet: Tetrodotoxin, apamin, iberiotoxin, and glibenclamide were purchased from Alomone Labs (Jerusalem, Israel), and Latoxan (Portes-lès-Valence, France).

Techniques: Inhibition

Journal: Toxins

Article Title: Gambierol Blocks a K+ Current Fraction without Affecting Catecholamine Release in Rat Fetal Adrenomedullary Cultured Chromaffin Cells

doi: 10.3390/toxins14040254

Figure Lengend Snippet: Action of gambierol on the voltage-dependence of K + current activation in rat fetal AMC cells. The current was measured at the end of 90 ms depolarizing steps from a holding potential of −70 mV, expressed as a percentage of its maximal value at +40 mV, and plotted as a function of membrane potential during depolarizing pulses, in the absence (AGI, in blue) and presence (in red) of 100 nM of gambierol. Data represent the mean ± SEM of 4 different experiments. The theoretical curves correspond to data point fits according to the Boltzmann equation, as described in Materials and Methods, with V 50% and k values of 14.25 mV and 8.07 mV −1 (R 2 = 0.9989), respectively, for AGI, and 8.93 mV and 10.17 mV − 1 (R 2 = 0.9997), respectively, in the presence of gambierol. AGI denotes the simultaneous perfusion of 400 nM apamin, 200 µM glibenclamide and 100 nM iberiotoxin.

Article Snippet: Tetrodotoxin, apamin, iberiotoxin, and glibenclamide were purchased from Alomone Labs (Jerusalem, Israel), and Latoxan (Portes-lès-Valence, France).

Techniques: Activation Assay

Journal: The Journal of Physiology

Article Title: Hypoxia activates ATP-dependent potassium channels in inspiratory neurones of neonatal mice

doi: 10.1111/j.1469-7793.1998.755bm.x

Figure Lengend Snippet: Pharmacological properties of K ATP channels with 75 pS conductance in Ca 2+ -free pipette solution Drugs were applied at the beginning of each recording and representative traces are shown at the times indicated. The dotted lines indicate the closed level and dashed lines indicate the open levels. A, time course of diazoxide action. Patch holding potential, 0 mV. B and C, two representative experiments of K ATP channel inhibition in inside-out patches by tolbutamide ( B ) and glibenclamide ( C ) at holding potentials of -25 and -40 mV, respectively. The data shown in B and C were obtained in patches that had not been subjected to any prior treatment.

Article Snippet: TTX and K+ channel blockers (tetraethylammonium (TEA), 4-aminopyridine, charybdotoxin, iberiotoxin, diazoxide, tolbutamide and glibenclamide) were purchased from Alomone Labs (Israel).

Techniques: Transferring, Inhibition

Journal: PLoS ONE

Article Title: Predominant Functional Expression of Kv1.3 by Activated Microglia of the Hippocampus after Status epilepticus

doi: 10.1371/journal.pone.0006770

Figure Lengend Snippet: Effects of α-dendrotoxin (A, 50 nM), agitoxin-2 (B, 10 and 50 nM) and margatoxin (C, 1 and 10 nM) on the leak subtracted current induced by a voltage step from −70 to +40 mV (black traces recorded in control, red traces after drug application). The leak conductances of the cells in A, B and C were 338, 862 and 332 pS, respectively. The graphs on the right represent the conductance, normalized to its maximum value, as a function of the membrane potential and its inhibition by α-dendrotoxin (A, n = 4, Dtx), agitoxin-2 (B, AgTx, n = 14 for 10 nM, n = 6 for 50 nM) and margatoxin (C, MgTX, n = 8 for 1 nM, n = 11 for 10 nM).

Article Snippet: Reagents Alpha-dendrotoxin, recombinant agitoxin-2 and recombinant margatoxin were purchased from Alomone labs (Jerusalem, Israel).

Techniques: Inhibition

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: P2X7 Receptor Mediates Spinal Microglia Activation of Visceral Hyperalgesia in a Rat Model of Chronic Pancreatitis

doi: 10.1016/j.jcmgh.2015.07.008

Figure Lengend Snippet: The abdominal reflex upon von Frey filament (VFF) stimuli in baseline showed no significant difference among the four groups (mixed-design two-way analysis of variance) in ( A ) brilliant blue G (BBG) treatment experiments ( P = .792); ( B ) BBG prevention experiments ( P = .401); and ( C ) P2X7R small-interfering RNA (siRNA) treatment experiments ( P = .247) .

Article Snippet: Furthermore, pharmacologic blockade of P2X7R via intraperitoneal administration of the selective inhibitors of P2X7R A-740003 (N -[1-[(E)-[(cyanoamino)-(quinolin-5-ylamino)methylidene]amino]-2,2-dimethylpropyl]-2-(3,4-dimethoxyphenyl)acetamide) and A-438079 (3-[[5-(2,3-dichlorophenyl)tetrazol-1-yl]methyl]pyridine) reduced tactile allodynia in three different rat models of neuropathic pain.

Techniques: Small Interfering RNA

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: P2X7 Receptor Mediates Spinal Microglia Activation of Visceral Hyperalgesia in a Rat Model of Chronic Pancreatitis

doi: 10.1016/j.jcmgh.2015.07.008

Figure Lengend Snippet: Brilliant blue G (BBG) treatment attenuated P2X7R expression in the spinal cord. ( A ) The expression level of P2X7R in the spinal cord was significantly increased in the trinitrobenzene sulfonic acid (TNBS)-treated rats (T + V) compared with the control rats (V + V). P2X7R expression was alleviated by intrathecal (IT) BBG treatment (T + B) ( P = .005, one-way analysis of variance [ANOVA]). * P

Article Snippet: Furthermore, pharmacologic blockade of P2X7R via intraperitoneal administration of the selective inhibitors of P2X7R A-740003 (N -[1-[(E)-[(cyanoamino)-(quinolin-5-ylamino)methylidene]amino]-2,2-dimethylpropyl]-2-(3,4-dimethoxyphenyl)acetamide) and A-438079 (3-[[5-(2,3-dichlorophenyl)tetrazol-1-yl]methyl]pyridine) reduced tactile allodynia in three different rat models of neuropathic pain.

Techniques: Expressing

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: P2X7 Receptor Mediates Spinal Microglia Activation of Visceral Hyperalgesia in a Rat Model of Chronic Pancreatitis

doi: 10.1016/j.jcmgh.2015.07.008

Figure Lengend Snippet: Brilliant blue G (BBG) administration prevents visceral hyperalgesia via the inhibition of P2X7R. ( A ) The mean abdominal reflexes in response to von Frey filament (VFF) stimulation in trinitrobenzene sulfonic acid (TNBS)-treated rats (V + T) were significantly higher than in the control rats (V + V). This effect was prevented by intrathecal (IT) BBG pretreatment (B + T) ( P

Article Snippet: Furthermore, pharmacologic blockade of P2X7R via intraperitoneal administration of the selective inhibitors of P2X7R A-740003 (N -[1-[(E)-[(cyanoamino)-(quinolin-5-ylamino)methylidene]amino]-2,2-dimethylpropyl]-2-(3,4-dimethoxyphenyl)acetamide) and A-438079 (3-[[5-(2,3-dichlorophenyl)tetrazol-1-yl]methyl]pyridine) reduced tactile allodynia in three different rat models of neuropathic pain.

Techniques: Inhibition

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: P2X7 Receptor Mediates Spinal Microglia Activation of Visceral Hyperalgesia in a Rat Model of Chronic Pancreatitis

doi: 10.1016/j.jcmgh.2015.07.008

Figure Lengend Snippet: P2X7R expression with OX-42-positive cells in dorsal horn of the spinal cord. In trinitrobenzene sulfonic acid (TNBS)-treated rats, P2X7R ( red ) staining colocalized with OX-42-positive cells (green) ( A, B ) but not with NeuN-positive cells ( green ) ( C, D ) or GFAP-positive cells (green) ( E, F ). Double immunofluorescence labeling for P2X7R and for cell-type specific markers: NeuN for neuron, GFAP for astrocyte, and OX42 for microglia. DAPI (4′,6-diamidino-2-phenylindole) staining identifies all cell nuclei ( blue ). Scale bar: 50 μm ( A, C, E ); 20 μm ( B, D, F ). The arrows indicate the colocalization of P2X7R expression with the microglia marker.

Article Snippet: Furthermore, pharmacologic blockade of P2X7R via intraperitoneal administration of the selective inhibitors of P2X7R A-740003 (N -[1-[(E)-[(cyanoamino)-(quinolin-5-ylamino)methylidene]amino]-2,2-dimethylpropyl]-2-(3,4-dimethoxyphenyl)acetamide) and A-438079 (3-[[5-(2,3-dichlorophenyl)tetrazol-1-yl]methyl]pyridine) reduced tactile allodynia in three different rat models of neuropathic pain.

Techniques: Expressing, Staining, Immunofluorescence, Labeling, Marker

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: P2X7 Receptor Mediates Spinal Microglia Activation of Visceral Hyperalgesia in a Rat Model of Chronic Pancreatitis

doi: 10.1016/j.jcmgh.2015.07.008

Figure Lengend Snippet: P2X7R small-interfering RNA (siRNA) treatment experiment. ( A , B , D ) Within-group analysis in the brilliant blue G (BBG) treatment experiments showed the abdominal reflexes upon von Frey filament (VFF) stimuli were similar before and after P2X7R siRNA/nontargeted siRNA treatment. ( C ) In the trinitrobenzene sulfonic acid (TNBS)-induced chronic pancreatitis (CP) rats, the abdominal reflexes upon VFF stimuli were significantly increased after nontargeted siRNA treatment. Analysis by two-way repeated measurement analysis of variance. ( A ) P = .391; ( B ) P = .069; ( C ) * P = .007; ( D ) P = .222.

Article Snippet: Furthermore, pharmacologic blockade of P2X7R via intraperitoneal administration of the selective inhibitors of P2X7R A-740003 (N -[1-[(E)-[(cyanoamino)-(quinolin-5-ylamino)methylidene]amino]-2,2-dimethylpropyl]-2-(3,4-dimethoxyphenyl)acetamide) and A-438079 (3-[[5-(2,3-dichlorophenyl)tetrazol-1-yl]methyl]pyridine) reduced tactile allodynia in three different rat models of neuropathic pain.

Techniques: Small Interfering RNA

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: P2X7 Receptor Mediates Spinal Microglia Activation of Visceral Hyperalgesia in a Rat Model of Chronic Pancreatitis

doi: 10.1016/j.jcmgh.2015.07.008

Figure Lengend Snippet: Pancreatic histology in chronic pancreatitis (CP) rats treated with vehicle ( A ), brilliant blue G (BBG) ( B ), and P2X7R small-interfering RNA (siRNA) ( C ). All groups of rats showed markedly inflammatory cells infiltration, large regions of acinar loss and periductular and intralobular fibrosis, and lost acinar tissue replaced with tubular structures. No significant morphologic changes could be observed after either BBG or siRNA treatment in CP rats. H E stain; scale bar: 100 μm.

Article Snippet: Furthermore, pharmacologic blockade of P2X7R via intraperitoneal administration of the selective inhibitors of P2X7R A-740003 (N -[1-[(E)-[(cyanoamino)-(quinolin-5-ylamino)methylidene]amino]-2,2-dimethylpropyl]-2-(3,4-dimethoxyphenyl)acetamide) and A-438079 (3-[[5-(2,3-dichlorophenyl)tetrazol-1-yl]methyl]pyridine) reduced tactile allodynia in three different rat models of neuropathic pain.

Techniques: Small Interfering RNA, Staining

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: P2X7 Receptor Mediates Spinal Microglia Activation of Visceral Hyperalgesia in a Rat Model of Chronic Pancreatitis

doi: 10.1016/j.jcmgh.2015.07.008

Figure Lengend Snippet: Increased P2X7R expression levels in the spinal cord after chronic pancreatitis (CP). ( A ) Immunohistochemistry of the spinal cord dorsal horn in vehicle-treated rats. ( B ) Three weeks after induction of CP, P2X7R expression was increased. ( C ) Representative Western blots of P2X7R expression in the thoracic spinal cord. The P2X7R expression level was significantly increased after CP. ( D ) The P2X7R mRNA expression level of was also significantly increased after CP. Scale bar = 100 μm in A and B . * P

Article Snippet: Furthermore, pharmacologic blockade of P2X7R via intraperitoneal administration of the selective inhibitors of P2X7R A-740003 (N -[1-[(E)-[(cyanoamino)-(quinolin-5-ylamino)methylidene]amino]-2,2-dimethylpropyl]-2-(3,4-dimethoxyphenyl)acetamide) and A-438079 (3-[[5-(2,3-dichlorophenyl)tetrazol-1-yl]methyl]pyridine) reduced tactile allodynia in three different rat models of neuropathic pain.

Techniques: Expressing, Immunohistochemistry, Western Blot

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: P2X7 Receptor Mediates Spinal Microglia Activation of Visceral Hyperalgesia in a Rat Model of Chronic Pancreatitis

doi: 10.1016/j.jcmgh.2015.07.008

Figure Lengend Snippet: Intrathecal (IT) administration of P2X7R-targeted small-interfering RNA (siRNA) ameliorated the chronic pancreatitis (CP)-related nocifensive behaviors. ( A ) The abdominal reflex in response to von Frey filament (VFF) stimulation in rats after intraductal infusion of TNBS and nontargeted siRNA treatment (T + N) was significantly increased compared with the control rats (V + N). This hyperalgesia was ameliorated by IT injection of P2X7R siRNA (T + KD) ( P

Article Snippet: Furthermore, pharmacologic blockade of P2X7R via intraperitoneal administration of the selective inhibitors of P2X7R A-740003 (N -[1-[(E)-[(cyanoamino)-(quinolin-5-ylamino)methylidene]amino]-2,2-dimethylpropyl]-2-(3,4-dimethoxyphenyl)acetamide) and A-438079 (3-[[5-(2,3-dichlorophenyl)tetrazol-1-yl]methyl]pyridine) reduced tactile allodynia in three different rat models of neuropathic pain.

Techniques: Small Interfering RNA, Injection