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1.
Fig. 3.

Fig. 3. From: Functional profiling of neurons through cellular neuropharmacology.

Size distributions of neurons responsive to select RA challenge compounds. Data are shown for only a few neuronal subclasses. The x axis is the same for all panels. Cell area is the area of a cross-section of the cell soma. (A) ACh only refers to neurons that responded to ACh but not to any other RA challenge compounds. (B) ACh + ATP + Cap refers to neurons that responded to ACh, ATP, and capsaicin but no other RA challenge compounds. (C) Menthol only refers to neurons that responded to menthol but no other RA challenge compound.

Russell W. Teichert, et al. Proc Natl Acad Sci U S A. 2012 January 31;109(5):1388-1395.
2.
Fig. 5.

Fig. 5. From: Functional profiling of neurons through cellular neuropharmacology.

Selected calcium-imaging traces from MP challenges. Each trace represents a single neuron's response. These data were obtained by using the MP challenge protocol exemplified by Fig. 4; x axis is the same for all traces, and the red color is for emphasis only. It highlights the different types of responses to each challenge compound. (A–E) Effects of 1 μM TTX, which was applied for 6 min starting at minute 23 (indicated by horizontal bars). (F–J) Effects of 25 mM TEA, which was applied for 6 min starting at minute 23 (bars). At the end of the experiment, 300 nM capsaicin was applied for 1 min at minute 57 (black circles).

Russell W. Teichert, et al. Proc Natl Acad Sci U S A. 2012 January 31;109(5):1388-1395.
3.
Fig. 1.

Fig. 1. From: Functional profiling of neurons through cellular neuropharmacology.

Images of dissociated mouse lumbar DRG neurons loaded with Fura-2-AM dye. A–D are images of the same field of view. Fluorescence images were acquired as described in Materials and Methods. (A) Bright-field image. (B) Fluorescence image acquired with 380-nm excitation and 510-nm emission filters. (C) Pseudocolored ratiometric calcium image obtained under control conditions (i.e., before depolarization). The color scale indicates that the resting cytoplasmic calcium concentration is relatively low (magenta and blue). (D) Pseudocolored ratiometric image obtained on depolarization of the neurons by 100 mM KCl. The color scale, same as the color scale in C, indicates that the cytoplasmic calcium concentration is relatively high in many cells (green, yellow, and red). (Scale bar: 30 μm.) Glial cells did not respond to 100 mM KCl with elevated cytoplasmic calcium, presumably because they lack voltage-gated calcium channels.

Russell W. Teichert, et al. Proc Natl Acad Sci U S A. 2012 January 31;109(5):1388-1395.
4.
Fig. 6.

Fig. 6. From: Functional profiling of neurons through cellular neuropharmacology.

Selected traces from large-diameter neurons (cell area > 600 μm2) illustrating the effects of voltage-gated K-channel blockers. Each trace represents a single neuron's response. The x axis is the same for all traces in a given panel. Challenge compounds were ACh, 1 mM acetylcholine; C, 300 nM capsaicin; K, 25 mM KCl; pl14a, 16 μM conopeptide pl14a; RIIIJ, 1 μM κM-conopeptide RIIIJ; and Dtx, 100 nM Dendrotoxin-K. Arrows and bars indicate when each challenge compound or KCl was applied; the latter was applied at 7-min intervals except as noted. The red color is for emphasis only. It highlights differences in responsiveness to the challenge compounds used. (A) ACh was applied (at minute 1) to identify responders (blue is for emphasis only), after which time the KCl pulses were given before and after application of κM-RIIIJ (bar). These neurons were resistant to capsaicin, which is typical for large-diameter DRG neurons. (B) KCl pulses presented before and after application of κM-RIIIJ (first bar) and conopeptide pl14a (second bar). These neurons were resistant to capsaicin. (C) KCl pulses applied before and after application of κM-RIIIJ (first bar) and Dtx-K (second bar).

Russell W. Teichert, et al. Proc Natl Acad Sci U S A. 2012 January 31;109(5):1388-1395.
5.
Fig. 4.

Fig. 4. From: Functional profiling of neurons through cellular neuropharmacology.

MP challenge protocol illustrated with calcium-imaging traces. Each trace represents a single neuron's response. Each peak in a trace is the response to a 25-mM KCl pulse applied at 7-min intervals. The duration of each KCl pulse (∼15 s) is indicated by the vertical bars (1–8). The first four KCl pulses (1–4) served as internal controls to monitor the variability in the elicited calcium signals. The last three KCl pulses (6–8) allowed us to monitor the reversibility of a compound's effects. The compound of interest was applied at minute 23 and remained in the bath for 6 min, which is indicated by the horizontal bar in Upper. The fifth KCl pulse, at minute 29 (peak 5), shows that the compound caused an amplification of the high [K+]o-elicited calcium signal, which was readily reversible (indicated by peaks 6–8). (Lower) Vehicle trials, in which only observation solution was applied to the bath at minute 23, served as external controls. The red color is for emphasis only. It highlights the difference between a response to a compound (Upper; compound trial) and a control without a compound (Lower; vehicle trial). At the end of the experiment, at minute 57, 300 nM capsaicin was applied for 1 min (black circles).

Russell W. Teichert, et al. Proc Natl Acad Sci U S A. 2012 January 31;109(5):1388-1395.
6.
Fig. 2.

Fig. 2. From: Functional profiling of neurons through cellular neuropharmacology.

Example calcium-imaging traces from RA challenges. Each trace is a single neuron's response to the challenge compounds indicated at the bottom of each panel. The x axis is the same for all traces in a given panel. The y axis (same for Figs. 4–6) is a relative measure of [Ca2+]i determined by the 340/380 nm excitation ratio described in Materials and Methods. Challenge compounds were ACh, 1 mM acetylcholine; ATP, 10 μM adenosine 5′-triphosphate; AITC, 200 μM allyl isothiocyanate; C, 300 nM capsaicin; H, 50 μM histamine; M, 200 μM menthol; K, 100 mM KCl (A–C) or 25 mM KCl (D); and TEA, 10 mM tetraethylammonium chloride. Arrows indicate when each compound was applied to the bath. (A–C) Typical sequence of RA challenges. Challenge compounds were applied at 5-min intervals for ∼15 s. ACh was applied two times to show reproducibility of responses. KCl (100 mM) was applied at the end of the series, and therefore, nonresponsive cells could be excluded from additional analysis. The maximum response to KCl was cropped for some traces, and therefore, heights of lesser peaks would be more evident. (C and D) ATP, menthol, and AITC were used to differentiate between neurons only sensitive to menthol and neurons sensitive to all three challenge compounds, which are highlighted in blue. (D) Responses from one neuron of each type are shown. Neurons were also depolarized by KCl (25 mM) pulses at 7-min intervals before and after application of 10 mM TEA (indicated by horizontal bar), a blocker of voltage-gated K channels, to compare TEA with KCl-elicited responses. The red color is for emphasis only. It highlights the different types of responses to TEA.

Russell W. Teichert, et al. Proc Natl Acad Sci U S A. 2012 January 31;109(5):1388-1395.

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