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. 2020 Mar 27;9:e55007. doi: 10.7554/eLife.55007

Figure 6. Unc-4 function is necessary in 7B neurons for the leg neuropil innervation .

Figure 6.

(A) An adult VNC containing MCFO clones showing two 7B neurons in the T2 segment; both extend projections (arrows) to the ipsilateral leg neuropil. (B) A subset of 7B neurons (magenta) in the T2 segment marked with SS20852-GAL4 driven syt-HA (yellow) showing synaptic outposts in the leg neuropil. (C, D) NB3-2 intersected reporter immortalization via ey-AD;dbxDBD driver used to visualize projections of 7B neurons in the control (C) and mutant (D) animals. Cartoons in the left panels schematized projections of the T2 7B neurons observed in confocal images. Transverse views (corresponding the yellow brackets) shown under the cartoons. In the mutant, ipsilateral (arrows) and contralateral (arrowheads) projections in the leg neuropils were reduced. Note that the numbers of T2 7B neurons marked with the reporter in control and mutant animals was the same (67 + - 6 and 68+/- 9 neurons in control and mutant animals, respectively; N = 7 clones for each). (E–G) A different immortalization technique involving FLP/FRT induced LexA:p65 without NB intersection used to mark 7B neurons in the VNC of control (E) and mutant (F,G) animals. ey-AD;dbxDBD expression was immortalized without NB intersection. Top panels show maximum z projections; bottom panels show the transverse view of the same region. As with the above manipulations, mutant 7B neurons failed to show the normal innervation of the leg neuropil (compare arrows in E and F). In some mutant animals, ipsilateral 7B projections extended posteriorly without innervating leg neuropils (arrows in G). (H–I) 7B MARCM clones in T2 segments of the control (H) and (I) unc-4 mutant animals shown. Both ipsilateral (arrows) and contralateral (arrowheads) projections into leg neuropils were reduced in the mutant clone (I). Control clone has 68 cells; unc-4 clone has 73 cells. (C–H) Maximum z projections were made of a subset of confocal slices to reveal neuronal processes in leg neuropils. n indicates the number of animals showing the phenotype. Leg neuropils are outlined with yellow dashed lines.