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. 2017 Sep 13;6:e29257. doi: 10.7554/eLife.29257

Figure 2. Axon fasciculation defects of casy-1 and rig-6 are distinguishable.

(A–D) The axons of the three neurons were individually labeled with wCherry (HOA; red), TagBFP (AVG; blue), and GFP (PHC; green) in wild type (A), casy-1(tm718) (B), rig-6(ok1589) (C), or casy-1;rig-6 double mutants (D). Axon fasciculation between each neuronal pair in the dashed box region and schematic of axon fasciculation are shown on the right. Arrowheads indicate the region where two axons are detached from each other. (E) Percentage of axon-axon contact between each neuronal pair in wild type or mutant animals (n = 40). Each dot represents individual animal. Red bar represents the median. (F) Developmental timing of axon extension of the three neurons in males. Each developmental stage was determined by the tail morphology of the male. Dashed lines indicate the posterior end of the intestine. (G) Axon length was measured from the point where the AVG axon makes a turn in the pre-anal ganglion (n = 15). Error bars are SEM. (H) Axon fasciculation of the three neurons in mock-, PHC-, HOA-, or AVG-ablated animal. Arrowheads indicate the region where HOA and AVG axons are detached from each other. (I) Percentage of axon-axon contact between each neuronal pair in mock-, PHC-, HOA-, or AVG-ablated animals. The number of animals analyzed is indicated. Scale bars, 20 μm. *p<0.05; **p<0.01; ***p<0.001; ns, not significant (by Mann-Whitney test). For the data and statistics, see Figure 2—source data 1.

Figure 2—source data 1. Source data for Figure 2 and Figure 2—figure supplement 2.
elife-29257-fig2-data1.xlsx (25.6KB, xlsx)
DOI: 10.7554/eLife.29257.010

Figure 2.

Figure 2—figure supplement 1. Comparison of PHCL-PHCR defasciculation in electron micrographs and fluorescence images.

Figure 2—figure supplement 1.

(A) Schematic of axon fasciculation between PHCL and PHCR neurons reconstructed from electron micrographs of an adult male N2Y (from section #13883 to #14249). Note that fasciculation of these axons is interrupted by single neurites of other neurons indicated. (B) Electron micrograph showing defasciculation of PHCL and PHCR axons with one-neurite distance (by a PVY neurite). (C) Fluorescence image of PHCL and PHCR axons (eat-4p11::gfp) was used to measure the length of fasciculation or defasciculation and this was summarized as a bar graph as in (A). Scale bar, 20 μm. (D) Summary of measurement of PHCL-PHCR fasciculation using fluorescence images obtained from 20 individual animals. The N2Y animal is shown for comparison.
Figure 2—figure supplement 2. Phenotypes of rig-6(gk438569) mutants.

Figure 2—figure supplement 2.

(A) Images of axon placement of HOA, AVG and PHC in rig-6(gk438569) mutants. Axon fasciculation between each neuronal pair in the dashed box region and schematic of axon fasciculation are shown on the right. Arrowheads indicate the region where two axons are detached from each other. Scale bar, 20 μm. (B) Percentage of axon-axon contact between each neuronal pair in rig-6(gk438569) mutant animals (n = 40). Each dot represents individual animal. Red bar represents the median. The data for wild type and rig-6(ok1589) mutants are identical to those in Figure 2 and are shown for comparison. *p<0.05; **p<0.01; ns, not significant (by Mann-Whitney test). For the data and statistics, see Figure 2—source data 1.