Figure 2. Cumulative serial ablation of Dbx1 neurons in preBötC-surface slices (A) and control slices whose surface exposes the ventral respiratory column, not preBötC (B).

(A and B) The x-axis is a timeline. The y-axis plots XII amplitude (normalized units, top) and respiratory period (bottom). The respiratory period axis is continuous (0–200 s) but plotted with two scales. Major ticks are separated by 10 s from 0 to 20 s (with unlabeled minor ticks at 5 s increments), and thereafter major ticks are plotted in 100 s divisions from 21 to 200 s (with unlabeled minor ticks at 50 s increments). The discontinuity in the y-axis stops at 20 s (lower portion) and starts at 21 s (upper portion). There is one data point for every individual respiratory period measured. The recording in A is no longer displayed after 6 min of XII quiescence. Substance P (SP) injection in A is displayed at higher sweep speed in Figure 5C. The recording in B is no longer displayed after 90 min of continuous stable XII output following the end of the ablation phase. Time calibrations in A and B are shown separately.

DOI: http://dx.doi.org/10.7554/eLife.03427.007

Figure 2—figure supplement 1. Cellular laser ablation and confirmation.

(A) The image acquired during maximum-intensity Ti:sapphire laser scanning of the target cell with a 560–615 nm band-pass filter, which indicates cell destruction. This image was acquired with higher digital magnification compared to panels B–D; scale bar is 2 µm. (B_1–2) Images of native tdTomato expression in Dbx1 preBötC neurons before (B₁) and after (B₂) a single cell laser ablation. The target cell (arrowhead) is visible pre-lesion but not in the post-lesion image. Neighboring (unlesioned) neurons are unaffected. Scale bar of 10 µm applies to all images in B–D. (C) Bright field images of the target cell (arrowhead) prior to laser lesion. (D_1–5) Images of the target cell post-lesion (arrowhead) at 5-µm increments in the z plane. The focal plane in C was normalized to z = 0 µm for relative comparison with panels D_1–5.

DOI: http://dx.doi.org/10.7554/eLife.03427.008

Figure 2—figure supplement 2. Cumulative tally of laser ablations for preBötC-surface slices (magenta) and control slices whose surface exposes the ventral respiratory column, not preBötC (cyan).

The total tally and the individual side tallies are shown for each preparation. Black bars show the mean. For preBötC-surface slices, the tally was always lower on the side that was being lesioned when the rhythm stopped because rhythm cessation halted the ablation sequence.

DOI: http://dx.doi.org/10.7554/eLife.03427.009