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. Author manuscript; available in PMC: 2012 Feb 11.
Published in final edited form as: Neuron. 2011 Aug 11;71(3):447–459. doi: 10.1016/j.neuron.2011.06.040

Figure 8. Rich, Rab6, and CadN function in a common pathway.

Figure 8

(A–F′) N-Cadherin is reduced in the lamina of rich and Rab6 mutants. Single optic slides of lamina of the fly eye at 24h APF stained with anti-CadN (red), 24B10 (blue), and GFP (green, mark the mutant cells). (A–C, A′–C′) show a side view of the lamina. (D–F, D′–F′) show a frontal view of the lamina. (A, A′, D, D′) control (y w eyFLP; Act>Gal4 UAS-SytGFP/UAS-SytGFP; FRT80B M (3), tub-Gal80/FRT 80B iso). (B, B′, E, E′) rich1 mutant (y w eyFLP; Act>Gal4 UAS-SytGFP/UAS-SytGFP; FRT80b M (3), tub-Gal80/FRT 80B rich1). (C, C′, F, F′) Rab6 mutant (y w eyFLP; FRT40A tub-Gal80/FRT40A Rab6D23D; tubGal4/UAS-SytGFP). (A′–F′) are red channels (CadN) of (A–F). (G–H′) CadN distribution is not affected in rich1 mutant cone cells. The pupal eye discs at 36APF were stained with anti-GFP (green) and anti-CadN (red) antibody. The mutant cells were marked with CD8GFP. The animal genotypes are: CTL (y w eyFLP; Act>UAS-SytGFP/UAS-CD8GFP; FRT80B M (3), tub-Gal80/FRT80B iso). rich1 (y w eyFLP; Act>UAS-SytGFP/UAS-CD8GFP; FRT80B M (3), tub-Gal80/FRT 80B rich1). (I) The R7 cell targeting patterns of rich1 and rich2 upon removal of one copy of CadN (y w UAS-SytGFP; FRT40A CadN405/CyO; FRT80B rich1/TM6b; and y w UAS-SytGFP; FRT40A CadN405/CyO; FRT80B rich2/TM6B). Compared with Figure 7(E) and (F), removal of one copy of CadN enhances the R7 targeting phenotypes of the hypomorphic allele rich2 but not the phenotypes of the null allele rich1, indicating Rich and CadN function in a same pathway to regulate R7 targeting. (J) The quantification of R7 targeting phenotypes of rich1 mutants (Figure 7E), rich2 mutants (Figure 7F) and mutants of rich1 and rich2 with one copy of CadN. Upon removal of one copy of CadN, 19.6 ± 2% (n=384) of R7 cells in rich1 and 19.0 ± 2 % (n=493) of R7 cells in rich2 mutants fail to target to the correct layer. (K) The CadN, rich1 double mutants have similar eye targeting phenotypes as CadN single mutants. The genotypes are: (CTL) y w eyFLP;; FRT80B cl ubi-GFP/FRT80B iso. (rich1): y w eyFLP;; FRT80B cl ubi-GFP/FRT80B rich1. (CadN): y w eyFLP; FRT40A CadNΔ14/FRT40A ubi-GFP. (CadN; rich1): y w eyFLP; FRT40A rich-gRE::VK37/FRT40A CadNΔ14; FRT80B rich1/3L6 deletion. (L) Quantification of mistargeted R7 of control flies (CTL): 0%, n=458; rich1 mutants, 18.9± 3.9%, n=450; CadN mutants, 24.6±2.2%, n= 244; CadN; rich1 mutants: 24.4±1.6%, n=237. (See also Figure S6–S8)