Electrophysiological comparison of left versus right stellate ganglia neurons

Figures

• 

  Fig. 1. Isolation, culturing and immunostaining of stellate ganglion neurons. A and B, Schematic drawing (A) and microscopic view (B) of the location of the stellate ganglia – adapted from [16]. C, Light microscopy showing the appearance of freshly isolated stellate ganglia neurons (Ci) and those neurons cultured for 5 days (Cii). D, Confocal microscopy of cultured stellate ganglia neurons stained for DAPI (Di, blue) and βIII tubulin (Dii, red). Scale bar is 25 μm. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

• 

  Fig. 2. Basic action potential (AP) characteristics of stellate ganglion neurons. A, Example of APs evoked by a 150 pA depolarising pulse (left panel) and the first derivative of these APs (right panel). The analysed parameters are schematically indicated. B, Dot plots of all AP parameters of 48 single neurons from 12 cell isolations. Each single cell recording is considered as an independent measure (see Methods section). C, Typical example of spontaneous APs (without current injection). D, Typical example of anodal break excitation in response to a 100 pA hyperpolarising pulse. Vrest = potential 5 ms before the first AP, APA = AP amplitude, MDP = maximal diastolic potential after the first AP, Vmax,dep = maximal AP upstroke velocity, Vmax,rep = maximal velocity of AP repolarisation, APD50 = AP duration at 50 % of repolarisation.

• 

  Fig. 3. Correlations between AP parameters. A, Two examples of APs with different Vrest evoked by a 150 pA depolarising pulse (top panel) and the first derivative of these APs (bottom panel). B to F, Linear relationships (red solid lines) of Vrest with APA (B), Vmax,dep (C), Vmax,rep (D), APD50 (E), and MDP (F) of 48 neurons. Each single cell recording is considered as an independent measure (see Methods section). R indicates the Pearson correlation coefficient. G, R values of all linear relationships between AP parameters. Green, pink and white horizontal bars indicate the strength of the correlations. *P < 0.05, #P < 0.001. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

• 

  Fig. 4. AP parameters of phasic and tonic neurons. A, Two examples of phasic firing evoked by a 150 pA, 500 ms depolarising pulse. B, Two examples of tonic firing evoked by a 150 pA, 500 ms depolarising pulse. C, Dot plot of the number of APs during the 150 pA, 500 ms depolarising pulse of 48 neurons, Each single cell recording is considered as an independent measure (see Methods section). D, Number of APs vs the depolarising pulses for phasic and tonic neurons. Data are average ± SEM. *P < 0.05 (Two-way RM ANOVA). E, Percentages of phasic and tonic neurons with spontaneous APs and anodal break excitation *P < 0.05 (Fisher exact test). F, Dot plots of the AP parameters in phasic (n = 31; closed circles) and tonic (n = 17, open circles) neurons. *P < 0.05 (unpaired t-test). ‘n’ indicates the number of cells; phasic firing was found in 6 out of 6 isolations, tonic firing in 5 out of 6 isolations.

• 

  Fig. 5. AP parameters in consecutive APs during a 150 pA depolarising step. A-F, Changes of cycle length (A), overshoot (B), MDP (C), Vmax,dep (D) Vmax,rep (E) and APD50 (F) during the first 20 APs of continuously firing tonic cells (n = 15). Each single cell recording is considered as an independent measure (see Methods section). Data are average ± SEM and is normalized to the AP of the 1st AP. *P < 0.05 1st vs consecutive APs (Friedman RM ANOVA on ranks).

• 

  Fig. 6. AP parameters of LSG and RSG neurons of female and male mice. A, Group design with female and male as main groups and LRS and RSG as subgroups. ‘n’ indicates the number of cells measured and each single cell recording is considered as an independent measure (see Methods section). The colours for the subgroups are used consistently in all panels. B, Dot plots of the AP parameters in female and male neurons (with LRS and RSG as subgroups). MDP in male neurons is significantly different from female neurons without subgroups differences. *P < 0.05 (nested t-test). C-E, Percentage of phasic firing neurons (C), spontaneous APs (D), and anodal breaks (E) of female and male LSG and RSG neurons. *P < 0.05 (multiple logistic regression). F, Group design with LSG and RSG as main groups and gender as subgroups. G, Dot plots of the AP parameters in LSG and RSG neurons (gender as subgroups).