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Figure 1. Activated µORs inhibit TRPM3-induced Ca2+ signals in somatosensory neurons and in an overexpression system. ; (a) DRG neurons were stimulated three times with PS (25 µM) and morphine (1 µM, red trace, n = 188 neurons, five recordings) or vehicle as control (black trace, n = 130 neurons, four recordings). (b) Instead of morphine, DAMGO (0.3 µM, blue trace, 47 neurons, three recordings) was applied (control, black trace, 59 neurons, three recordings). Images of cells isolated from DRGs as used in these experiments are presented in Figure 1—figure supplement 5, example traces of individual cells in Figure 1—figure supplement 6a. (c) Quantification of the inhibition measured in (a) and (b); each dot-like symbol next to the bars represents the inhibition measured and calculated for a single, individual cell. Control experiments showing that the recorded Ca2+ signals are TRPM3-dependent are presented in Figure 1—figure supplement 1. Similar data with two chemically distinct µOR agonists are shown in Figure 1—figure supplement 2. (d) Inhibition by 0.3 µM DAMGO and recovery in DRG neurons stimulated with 25 µM PS, 0.3 µM DAMGO as indicated (n = 111 neurons, seven recordings). (e) The opioid receptor antagonist naloxone (5 µM) did not have an effect on its own (n = 412 neurons, 14 recordings). (f) Naloxone blocked the inhibitory effect of DAMGO on TRPM3 (n = 364 neurons, 15 recordings). Example traces of individual cells from these experiments are shown in Figure 1—figure supplement 6b. (g) Statistical analysis of the recordings shown in panels (d–f). The inhibition caused by DAMGO or naloxone was evaluated. The symbols on the right side of the bars represent inhibition measured in individual cells. (h) In HEK cells overexpressing TRPM3 and µORs, morphine (1 µM) inhibited Ca2+ signals evoked by 25 µM PS (orange trace, n = 115 cells, four recordings), but not in control cells expressing only TRPM3 (green trace, n = 97 cells, same four recordings). (i) DAMGO (0.3 µM) inhibits TRPM3-dependent Ca2+ signals induced by 50 µM nifedipine (orange trace: n = 58 cells, green trace: n = 75 cells, both from three recordings). (j) DAMGO (0.3 µM) inhibits TRPM3-dependent Ca2+ signals induced by heat (upper panel). Orange trace (n = 59 cells from four recordings) is the average of cells overexpressing TRPM3 and µORs. The blue trace (n = 89 cells, three recordings) represents control measurements from HEK cells expressing only µORs; there, TRPM3-independent heat-evoked Ca2+ signals are not inhibited by µOR activation. The lower panel shows the time course of the applied temperature. (k) Exemplary patch-clamp recording of a HEK cell overexpressing TRPM3 (activated by 25 µM PS) and µORs (activated by 0.3 µM DAMGO). Traces were obtained at +80 and −80 mV from voltage ramps. (l) I/V-curves measured at time points indicated in (k). (m) Statistical analysis of the baseline-subtracted current densities (n = 15 cells). A graphical representation of the recorded values for each cell is given in Figure 1—figure supplement 3. A dose response curve for the DAMGO-induced inhibition of TRPM3 currents is shown in Figure 1—figure supplement 4.

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Academic Journal

Reference temperature (°K) Shift factor

  • Source: http://proceedings.asmedigitalcollection.asme.org/data/Conferences/ASMEP/83707/V005T12A008-87-GT-81.pdf.

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T Temperature, K

  • Source: http://techreports.larc.nasa.gov/ltrs/PDF/jtht-8-2-94.pdf.

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