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. 2013 Mar 5;2:e00337. doi: 10.7554/eLife.00337

Figure 3. Quantitative analysis of the DCN soma and axon.

(A) A typical DCN axonal pattern showing GFP-labeled neurites (green) and LacZ labeled soma (red). Scale bar: 20 µm. (B) Analysis of interaxonal distances of DCN medulla axons along the D-V and A-P axes. Medulla axons are less dense medially and more dense dorsally and ventrally (n = 16). (C) Projection of a typical DCN cluster with sparse soma dorsally and ventrally and a high soma density medially. Scale bar: 5 µm. (D) 3D stem plot of the DCN cluster shown in (C) displays lines extending form P-D and A-P axis while the 3D position is marked with a filled circle—red indicates ventral cells, blue shows medial, and magenta shows dorsal cells. Vertical lines along the P-D and A-P axes form a representation of the density of the DCNs. (E) Ratio of the number of DCNs and their medulla axons along the D-V axis (n = 16). While dorsally and ventrally 1–2 neurons generate one medulla axon, medially 4–8 neurons produce one medulla axon. (F) Schematic representation of the sub-cluster model: a DCN cluster is divided into several overlapping subsets of neighboring neurons. Each subset provides one medulla axon which leads to pattern of spaced single axons in the medulla. A-P: anterior–posterior; DCN: dorsal cluster neurons; D-V: dorsal–ventral; P-D: proximal–distal.

DOI: http://dx.doi.org/10.7554/eLife.00337.006

Figure 3.

Figure 3—figure supplement 1. Counting DCN soma and medulla axons.

Figure 3—figure supplement 1.

Technical schematic of the labeling of DCN soma (red) and medulla axons (green) with Cartesian 3D coordinates (yellow dots for DCNs, red dots for medulla axons). DCN: dorsal cluster neurons.
DOI: http://dx.doi.org/10.7554/eLife.00337.007
Figure 3—figure supplement 2. Variability of the DCN cluster shape induces robust wiring pattern.

Figure 3—figure supplement 2.

(Result of a DCN density analysis and their medulla axons in two different genetic backgrounds (first row) n = 20 vs (second row) n = 14. First row, typical pattern of the DCNs and their medulla axons. Scale bar: 20 µm. Density chart of the medulla axons. A typical dorsal cluster of each genetic background. Scale bar: 5 µm. Fourth column, 3D reconstruction of third column. Fifth column, charts representing the ratio between neurons and their medulla axons. The axonal pattern was divided in 10 and 11 subunits according to the genetic background and the axonal pattern. These subunits were transferred to the DCN to count the number of neurons per subunit. DCN: dorsal cluster neurons.
DOI: http://dx.doi.org/10.7554/eLife.00337.008