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Laser ablation of Dbx1 neurons in the pre-Bötzinger complex stops inspiratory rhythm and impairs output in neonatal mice.

Wang X, Hayes JA, Revill AL, Song H, Kottick A, Vann NC, LaMar MD, Picardo MC, Akins VT, Funk GD, Del Negro CA - Elife (2014)

Bottom Line: To understand the neural origins of rhythmic behavior one must characterize the central pattern generator circuit and quantify the population size needed to sustain functionality.Breathing-related interneurons of the brainstem pre-Bötzinger complex (preBötC) that putatively comprise the core respiratory rhythm generator in mammals are derived from Dbx1-expressing precursors.These results demonstrate that a single canonical interneuron class generates respiratory rhythm and contributes in a premotor capacity, whereas these functions are normally attributed to discrete populations.

View Article: PubMed Central - PubMed

Affiliation: Department of Applied Science, The College of William and Mary, Williamsburg, United States.

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Average number of Dbx1 neurons detected at each acquisition depth from z = 0 (surface) to z = −80 µm in preBötC-surface slices and control slices with the ventral respiratory column (VRC) exposed at the slice surface.The number of Dbx1 neurons detected per focal plane per side (in 10-µm increments of the focal plane) is shown individually for each individual experiment (graygray unfilled circles) along with the mean ±SD for all experiments (black unfilled squares with black lines showing SD).DOI:http://dx.doi.org/10.7554/eLife.03427.006
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fig1s3: Average number of Dbx1 neurons detected at each acquisition depth from z = 0 (surface) to z = −80 µm in preBötC-surface slices and control slices with the ventral respiratory column (VRC) exposed at the slice surface.The number of Dbx1 neurons detected per focal plane per side (in 10-µm increments of the focal plane) is shown individually for each individual experiment (graygray unfilled circles) along with the mean ±SD for all experiments (black unfilled squares with black lines showing SD).DOI:http://dx.doi.org/10.7554/eLife.03427.006

Mentions: In the subsequent ‘detection phase’, a visible wavelength laser scanned the domain and an iterative threshold-crossing algorithm analyzed the image to then draw regions of interest (ROIs) for putative cell targets based on fluorescence brightness. Potential targets were evaluated on the basis of shape to differentiate cell bodies from auto-fluorescent debris, and to reject the fluorescence from dendrites and neuropil whose somata were detectable in adjacent focal planes (Wang et al., 2013) (Figure 1—figure supplement 2). The map of ROIs for validated cell targets was retained at each focal plane (Figure 1C—figure supplements 1B and 2, red ROIs). Potential targets that did not meet these criteria were discarded (but displayed for demonstration purposes in Figure 1C and Figure 1—figure supplements 1B and 2, blue ROIs). Target detection was repeated at 10-µm increments through the z-axis and the final three-dimensional map of targets was stored in memory (Figure 1D). Typically, we detected 26–50 Dbx1 neurons per focal plane per side (Figure 1—figure supplement 3) for a total average number of 705 targets in the preBötC (SD 119, SEM 59, range: 548 to 802, n = 8 slices).


Laser ablation of Dbx1 neurons in the pre-Bötzinger complex stops inspiratory rhythm and impairs output in neonatal mice.

Wang X, Hayes JA, Revill AL, Song H, Kottick A, Vann NC, LaMar MD, Picardo MC, Akins VT, Funk GD, Del Negro CA - Elife (2014)

Average number of Dbx1 neurons detected at each acquisition depth from z = 0 (surface) to z = −80 µm in preBötC-surface slices and control slices with the ventral respiratory column (VRC) exposed at the slice surface.The number of Dbx1 neurons detected per focal plane per side (in 10-µm increments of the focal plane) is shown individually for each individual experiment (graygray unfilled circles) along with the mean ±SD for all experiments (black unfilled squares with black lines showing SD).DOI:http://dx.doi.org/10.7554/eLife.03427.006
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4129438&req=5

fig1s3: Average number of Dbx1 neurons detected at each acquisition depth from z = 0 (surface) to z = −80 µm in preBötC-surface slices and control slices with the ventral respiratory column (VRC) exposed at the slice surface.The number of Dbx1 neurons detected per focal plane per side (in 10-µm increments of the focal plane) is shown individually for each individual experiment (graygray unfilled circles) along with the mean ±SD for all experiments (black unfilled squares with black lines showing SD).DOI:http://dx.doi.org/10.7554/eLife.03427.006
Mentions: In the subsequent ‘detection phase’, a visible wavelength laser scanned the domain and an iterative threshold-crossing algorithm analyzed the image to then draw regions of interest (ROIs) for putative cell targets based on fluorescence brightness. Potential targets were evaluated on the basis of shape to differentiate cell bodies from auto-fluorescent debris, and to reject the fluorescence from dendrites and neuropil whose somata were detectable in adjacent focal planes (Wang et al., 2013) (Figure 1—figure supplement 2). The map of ROIs for validated cell targets was retained at each focal plane (Figure 1C—figure supplements 1B and 2, red ROIs). Potential targets that did not meet these criteria were discarded (but displayed for demonstration purposes in Figure 1C and Figure 1—figure supplements 1B and 2, blue ROIs). Target detection was repeated at 10-µm increments through the z-axis and the final three-dimensional map of targets was stored in memory (Figure 1D). Typically, we detected 26–50 Dbx1 neurons per focal plane per side (Figure 1—figure supplement 3) for a total average number of 705 targets in the preBötC (SD 119, SEM 59, range: 548 to 802, n = 8 slices).

Bottom Line: To understand the neural origins of rhythmic behavior one must characterize the central pattern generator circuit and quantify the population size needed to sustain functionality.Breathing-related interneurons of the brainstem pre-Bötzinger complex (preBötC) that putatively comprise the core respiratory rhythm generator in mammals are derived from Dbx1-expressing precursors.These results demonstrate that a single canonical interneuron class generates respiratory rhythm and contributes in a premotor capacity, whereas these functions are normally attributed to discrete populations.

View Article: PubMed Central - PubMed

Affiliation: Department of Applied Science, The College of William and Mary, Williamsburg, United States.

Show MeSH
Related in: MedlinePlus