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. 2012 Oct 1;26(19):2206–2221. doi: 10.1101/gad.193409.112

Figure 1.

Figure 1.

MIG-10B localizes to AIY presynaptic regions in response to Netrin. (A) Confocal micrograph of cytoplasmic GFP expressed cell-specifically in AIY, depicting the morphology of AIY. (B–F) Synaptic vesicle pattern in maximal projection confocal micrographs of GFP∷RAB-3 in AIY in wild-type (B), mig-10(ct41) (D), and mig-10(ok2499) (E) mutant animals and schematics (C,F). The schematic diagram of wild-type AIY (shown in C) indicates neurite zones. Zone 1 is asynaptic and projects anteriorly from the cell body. AIY forms synapses onto RIA, AIZ, and RIB in zone 2. Zone 3 is within the nerve ring and is the distal portion of the neurite after the neurite turns dorsally (Colon-Ramos et al. 2007). Bar: B, 5 μm (applies to A,B,D,E). (G) Diagram of the mig-10 gene, with exons as boxes and introns as lines. The first three schematics show the relative positions of the three MIG-10 isoforms—MIG-10A, MIG-10B, and MIG-10C—drawn to scale. Shared exons code for conserved domains. Cartoon diagram of the conserved protein domains in the top right corner. Ras association (RA), plekstrin homology (PH), proline-rich (Prl), and FPPP (FP) (Holt and Daly 2005). The fourth and fifth schematics include a diagram of the rescuing fosmids, drawn to scale and aligned with the genetic regions of mig-10 and the isoforms. Modified image adapted with permission from WormBase (http://www.wormbase.org). (H–J) Confocal micrographs showing the cell-specific subcellular localization of MIG-10A∷GFP (H), MIG-10B∷GFP (I), and MIG-10C∷GFP (J). Note that MIG-10A and MIG-10C signal is diffuse throughout the neurite, including asynaptic regions (bracket) (H,J), while MIG-10B is enriched in presynaptic regions (I, arrows). (K,L) Confocal micrographs showing MIG-10B∷GFP subcellular localization in AIY in unc-6(ev400) (K) and unc-40(e271) (L) mutant animals. (I) Note that MIG-10B∷GFP is less enriched in zone 2 in the two mutant backgrounds, as compared with wild-type animals. (M–O) Confocal micrographs of zones 2 and 3 of a transgenic animal expressing MIG-10B∷GFP (M) and mCh∷RAB-3 (N) in AIY. Merge is shown in O. Synaptic regions are expanded in these images to allow evaluation of partial colocalization (arrows). In all images, the asterisk (*) represents the location of the cell body, and the dashed box encloses zone 2. Bar: H, 5 μm (applies to H–L); M, 5 μm (applies to M–O).

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