Functional distribution of Ca-coupled P2 purinergic receptors among adrenergic and noradrenergic bovine adrenal chromaffin cells-3

<p><b>Copyright information:</b></p><p>Taken from "Functional distribution of Ca-coupled P2 purinergic receptors among adrenergic and noradrenergic bovine adrenal chromaffin cells"</p><p>http://www.biomedcentral.com/1471-2202/8/39</p><p>BMC Neuroscience 2007;8():39-39.</p><p>Published online 14 Jun 2007</p><p>PMCID:PMC1906789.</p><p></p>acellular calcium (B) and during stimulation in the virtual absence of extracellular calcium (C, "UTP+EGTA"). At the end of each experiment cells were stimulated with 10 μM nicotine. Cells were allowed to rest for 10 min between consecutive stimulations. The fura-2 fluorescence ratio F/Fwas determined for each cell in a field on a pixel-by-pixel basis. Images were coded in pseudocolor to show differences in the F/Fratio. The images corresponding to UTP stimulation (B and C) were captured ~5 s after the UTP challenges (i.e. at the response peak in presence of extracellular calcium); . Pseudocolor image of PNMT/TH immunofluorescence ratio (labeling index) and corresponding scale. Intensely colored red and green cells are adrenergic and noradrenergic cells, respectively. Scale bar = 50 μm; . Time courses of changes in F/Ffluorescence ratio for two AD-cells (cells 1 and 2, also depicted in A-D) and two NA-cells (cells 3 and 4). The lines denote superfusions with UTP or nicotine in the presence or virtual absence (middle traces) of extracellular calcium

Selective stimulation of catecholamine release from bovine adrenal chromaffin cells by an ionotropic purinergic receptor sensitive to 2-methylthio ATP-2

<p><b>Copyright information:</b></p><p>Taken from "Selective stimulation of catecholamine release from bovine adrenal chromaffin cells by an ionotropic purinergic receptor sensitive to 2-methylthio ATP"</p><p>http://www.biomedcentral.com/1471-2202/8/41</p><p>BMC Neuroscience 2007;8():41-41.</p><p>Published online 20 Jun 2007</p><p>PMCID:PMC1906790.</p><p></p>owed to rest for ~10 min between successive applications. Nominal agonist concentrations (in μM) are indicated beneath each trace. Nominal concentrations of test agents are used throughout this legend. Actual concentrations at the cell bed differ from nominal concentrations by a factor of 1.64 (see Methods). Concentrations along the X axis in Fig. 3B are actual concentrations at the cell bed; . Dose-response curves of catecholamine release evoked by 2-MeSATP and ATP (circles and squares, respectively). Data from the experiment depicted in A and two similar experiments. Also depicted are data for UTP (100 μM; filled square) as well as for 2-MeSATP stimulations in presence of 50 μM PPADS (diamond), in the virtual absence of extracellular Ca(EGTA-containing solution, inverted triangle) and in absence of extracellular Na(Nareplacement for NMG, triangle) (n = 3 experiments for each condition)

Functional distribution of Ca-coupled P2 purinergic receptors among adrenergic and noradrenergic bovine adrenal chromaffin cells-2

<p><b>Copyright information:</b></p><p>Taken from "Functional distribution of Ca-coupled P2 purinergic receptors among adrenergic and noradrenergic bovine adrenal chromaffin cells"</p><p>http://www.biomedcentral.com/1471-2202/8/39</p><p>BMC Neuroscience 2007;8():39-39.</p><p>Published online 14 Jun 2007</p><p>PMCID:PMC1906789.</p><p></p>lumn in each histogram represents unresponsive cells (ΔR < 0.2). "ATP + Ca": responses obtained in presence of extracellular calcium; "ATP + EGTA": responses obtained in the virtual absence of extracellular calcium. , leftmost histograms: AD-cells; , rightmost histograms: NA-cells. A lognormal distribution function was fitted to each histogram, not taking into account unresponsive cells. The horizontal bars represent mean ΔR ± S.D

Functional distribution of Ca-coupled P2 purinergic receptors among adrenergic and noradrenergic bovine adrenal chromaffin cells-4

<p><b>Copyright information:</b></p><p>Taken from "Functional distribution of Ca-coupled P2 purinergic receptors among adrenergic and noradrenergic bovine adrenal chromaffin cells"</p><p>http://www.biomedcentral.com/1471-2202/8/39</p><p>BMC Neuroscience 2007;8():39-39.</p><p>Published online 14 Jun 2007</p><p>PMCID:PMC1906789.</p><p></p>olumn in each histogram represents unresponsive cells (ΔR < 0.2). "UTP + Ca": responses obtained in presence of extracellular calcium; "UTP + EGTA": responses obtained in the virtual absence of extracellular calcium. , leftmost histograms: AD-cells; , rightmost histograms: NA-cells. A lognormal distribution function was fitted to each histogram, not taking into account unresponsive cells. The horizontal bars represent mean ΔR ± S.D

Selective stimulation of catecholamine release from bovine adrenal chromaffin cells by an ionotropic purinergic receptor sensitive to 2-methylthio ATP-0

<p><b>Copyright information:</b></p><p>Taken from "Selective stimulation of catecholamine release from bovine adrenal chromaffin cells by an ionotropic purinergic receptor sensitive to 2-methylthio ATP"</p><p>http://www.biomedcentral.com/1471-2202/8/41</p><p>BMC Neuroscience 2007;8():41-41.</p><p>Published online 20 Jun 2007</p><p>PMCID:PMC1906790.</p><p></p>M UTP (D). At the end of each experiment cells were stimulated with 10 μM DMPP. Cells were allowed to rest for ~10 min between consecutive stimulations. The fura-2 fluorescence ratio F/Fwas determined for each cell in a field on a pixel-by-pixel basis. Images were coded in pseudocolor to show differences in the F/Fratio. The images corresponding to agonist stimulation (B-D) were captured at the response peaks. Calibration bar: 50 μm; . Time courses of changes in F/Ffluorescence ratio for a 2-MeSATP-sensitive/UTP-insensitive cell (cell 1), an UTP-sensitive/2-MeSATP-insensitive cell (cell 2), a cell displaying a mixed response (cell 3) and an ATP-insensitive cell (cell 4). The lines denote superfusions with DMPP or purinergic agonists. Some of the peak responses to DMPP were truncated for scaling reasons; . Frequency distribution histogram of calcium responses. Changes in ΔR = F/Fwere determined from the experiment depicted in A-D and 2 similar experiments (n = 234 cells). The column at the origin represents cells that did not respond to either UTP or 2-MeSATP (ΔR < 0.5, n = 137 cells). This column was truncated for scaling reasons. Columns in orange: cells responding to 2-MeSATP only; columns in green: cells responding to UTP only; columns in yellow: cells exhibiting mixed responses

Selective stimulation of catecholamine release from bovine adrenal chromaffin cells by an ionotropic purinergic receptor sensitive to 2-methylthio ATP-1

<p><b>Copyright information:</b></p><p>Taken from "Selective stimulation of catecholamine release from bovine adrenal chromaffin cells by an ionotropic purinergic receptor sensitive to 2-methylthio ATP"</p><p>http://www.biomedcentral.com/1471-2202/8/41</p><p>BMC Neuroscience 2007;8():41-41.</p><p>Published online 20 Jun 2007</p><p>PMCID:PMC1906790.</p><p></p>ation in the virtual absence of extracellular calcium ("2-MeSATP +EGTA", B); . The cells were subsequently superfused with a Ca-containing solution and challenged with 2-MeSATP, in the absence (C) or presence of 50 μM PPADS ("2-MeSATP +PPADS", D). Calibration bar: 50 μm. More in the legend to Fig. 1(A-D); . Time courses of changes in F/Ffluorescence ratio for two representative cells (cells 1 and 2, also depicted in A-D). The lines denote superfusions with 2-MeSATP, PPADS or EGTA-containing solution. The responses are representative of data from 3 experiments

Selective stimulation of catecholamine release from bovine adrenal chromaffin cells by an ionotropic purinergic receptor sensitive to 2-methylthio ATP-4

<p><b>Copyright information:</b></p><p>Taken from "Selective stimulation of catecholamine release from bovine adrenal chromaffin cells by an ionotropic purinergic receptor sensitive to 2-methylthio ATP"</p><p>http://www.biomedcentral.com/1471-2202/8/41</p><p>BMC Neuroscience 2007;8():41-41.</p><p>Published online 20 Jun 2007</p><p>PMCID:PMC1906790.</p><p></p>ation in the virtual absence of extracellular calcium ("2-MeSATP +EGTA", B); . The cells were subsequently superfused with a Ca-containing solution and challenged with 2-MeSATP, in the absence (C) or presence of 50 μM PPADS ("2-MeSATP +PPADS", D). Calibration bar: 50 μm. More in the legend to Fig. 1(A-D); . Time courses of changes in F/Ffluorescence ratio for two representative cells (cells 1 and 2, also depicted in A-D). The lines denote superfusions with 2-MeSATP, PPADS or EGTA-containing solution. The responses are representative of data from 3 experiments

Functional distribution of Ca-coupled P2 purinergic receptors among adrenergic and noradrenergic bovine adrenal chromaffin cells-1

<p><b>Copyright information:</b></p><p>Taken from "Functional distribution of Ca-coupled P2 purinergic receptors among adrenergic and noradrenergic bovine adrenal chromaffin cells"</p><p>http://www.biomedcentral.com/1471-2202/8/39</p><p>BMC Neuroscience 2007;8():39-39.</p><p>Published online 14 Jun 2007</p><p>PMCID:PMC1906789.</p><p></p>acellular calcium (B) and during stimulation in the virtual absence of extracellular calcium (C, "ATP+EGTA"). At the end of each experiment cells were stimulated with 10 μM DMPP. Cells were allowed to rest for 10 min between consecutive stimulations. The fura-2 fluorescence ratio F/Fwas determined for each cell in a field on a pixel-by-pixel basis. Images were coded in pseudocolor to show differences in the F/Fratio. The images corresponding to ATP stimulation (B and C) were captured ~5 s after the ATP challenges (i.e. at the response peak in presence of extracellular calcium); . Pseudocolor image of PNMT/TH immunofluorescence ratio (labeling index) and corresponding scale. Intensely colored red and green cells are adrenergic and noradrenergic cells, respectively. Scale bar = 50 μm. . Time courses of changes in F/Ffluorescence ratio for three AD-cells (cells 1, 2 and 5, also depicted in A-D) and two NA-cells (cells 3 and 4). The lines denote superfusions with ATP or DMPP in the presence or virtual absence (middle traces) of extracellular calcium

Functional distribution of Ca-coupled P2 purinergic receptors among adrenergic and noradrenergic bovine adrenal chromaffin cells-6

<p><b>Copyright information:</b></p><p>Taken from "Functional distribution of Ca-coupled P2 purinergic receptors among adrenergic and noradrenergic bovine adrenal chromaffin cells"</p><p>http://www.biomedcentral.com/1471-2202/8/39</p><p>BMC Neuroscience 2007;8():39-39.</p><p>Published online 14 Jun 2007</p><p>PMCID:PMC1906789.</p><p></p>energic cells); . Pseudocolor image of PNMT/TH immunofluorescence ratio (labeling index) and corresponding scale (right). Yellow corresponds to an index of ~98, the center of the mis-identification region (84–112) indicated by the black box on the scale; . Frequency distribution histogram for all cells analyzed in this study (number of cells . labeling index, measured as the averaged index over the region covered by the cell). Cells with a labeling index above the upper threshold (toward red) were considered PNMT(adrenergic) cells. Conversely, cells with an index below the lower threshold were considered TH/PNMT(noradrenergic) cells. Controls without primary antibodies showed a very faint, almost indiscernible fluorescence, in photographs taken using the same exposure times. Scale bar = 50 μm

Functional distribution of Ca-coupled P2 purinergic receptors among adrenergic and noradrenergic bovine adrenal chromaffin cells-5

<p><b>Copyright information:</b></p><p>Taken from "Functional distribution of Ca-coupled P2 purinergic receptors among adrenergic and noradrenergic bovine adrenal chromaffin cells"</p><p>http://www.biomedcentral.com/1471-2202/8/39</p><p>BMC Neuroscience 2007;8():39-39.</p><p>Published online 14 Jun 2007</p><p>PMCID:PMC1906789.</p><p></p>lar calcium. Panel A depicts cells prior to stimulation (control). Cells were allowed to rest for 30 min between consecutive stimulations. The SBFI fluorescence ratio F/Fwas determined for each cell in a field on a pixel-by-pixel basis. Images were coded in pseudocolor to show differences in the F/Fratio. The images corresponding to ATP and UTP stimulation (B and C) were captured ~30 s after challenges (response peaks); . Pseudocolor image of PNMT/TH immunofluorescence ratio (labeling index) and corresponding scale. Intensely colored red and green cells are adrenergic and noradrenergic cells, respectively. Scale bar = 50 μm. . Time course of changes in F/Ffluorescence ratio for an NA-cell (depicted by an arrow in A-D). The lines denote superfusions with ATP and UTP; . Frequency distribution histograms of sodium responses to ATP. Changes in ΔR= F/Fwere determined from the experiment depicted in A-B and 11 similar experiments (n = 749 cells). The first column in each histogram represents ATP-unresponsive cells (ΔR < 0.05). Leftmost histogram: AD-cells; rightmost histogram: NA-cells. A lognormal distribution function was fitted to each histogram, not taking into account unresponsive cells. The horizontal bars represent mean ΔR ± S.D