Introduction Nübler-Jung et al., 1987 Klein and Mlodzik, 2005 Drosophila Wallingford et al., 2002 Drosophila Fig. 1 Tree et al., 2002a; Klein and Mlodzik, 2005; Strutt and Strutt, 2005 Adler et al., 1998; Zeidler et al., 2000; Strutt and Strutt, 2002; Ma et al., 2003 widerborst Hannus et al., 2002 fz Shulman et al., 1998 Drosophila Strutt, 2001 Axelrod, 2001; Das et al., 2004 Bastock et al., 2003; Tree et al., 2002b Chae et al., 1999; Usui et al., 1999 Fig. 1 brokenheart Katanaev et al., 2005 fz ds Strutt and Strutt, 2002; Matakatsu and Blair, 2004 Lawrence et al., 2004; Amonlirdviman et al., 2005 Adler et al., 2000; Ma et al., 2003; Lawrence et al., 2004 Strutt and Strutt, 2002 Park et al., 1994; Zheng et al., 1995; Adler et al., 1997 Tree et al., 2002b; Lawrence et al., 2004; Amonlirdviman et al., 2005; Klein and Mlodzik, 2005; Le Garrec et al., 2006 Wong and Adler, 1993; Strutt, 2001 Strutt and Strutt, 2002 fz Classen et al., 2005 Strutt and Strutt, 2002; Lawrence et al., 2004 Tree et al., 2002b; Amonlirdviman et al., 2005 Tree et al., 2002b; Jenny et al., 2005 Bastock et al., 2003; Jenny et al., 2005 Das et al., 2004 Das et al., 2004 In this manuscript, we address three key issues: First, which components of the core act together with Fz during the different planar polarity patterning processes? Second, are the spatial relationships seen during the later phase of asymmetric localisation also relevant during the phase of cell–cell communication and local coordination of polarity? Third, is asymmetric core protein localisation absolutely required for planar polarity patterning? Materials and methods Fly strains and genetics Strutt and Strutt, 2002 Actin fz-EYFP Actin stbm-EYFP Strutt, 2001; Strutt et al., 2002 Actin dsh-ECFP Actin fmi-FLAG Actin pk pk sev-stbm sev-pk sple Strutt and Strutt, 2002 pk Gubb et al., 1999 fz Arm-fz-EGFP Strutt, 2001 stbm/Vang Actin-stbm-EYFP fz;stbm FRT42 stbm 6 Arm-fz-EGFP fz stbm 6 Arm-fz-EGFP Ubx-FLP Exact genotypes used are as follows: Figure 1 stbm/Vang w hsFLP1; stbm 6 /stbm Vang-A3 ; Act-FRT-polyA-FRT-stbm-EYFP/+ pk pk-sple w hsFLP1; pk pk-sple-13 /pk pk-sple-13 ; Act-FRT-polyA-FRT-pk/+ w; FRT42 stbm 6 /FRT42 P[w + ] stbm Vang-A3 stbm/Vang sev-stbm w; FRT42 stbm 6 /FRT42 P[w + ]stbm Vang-A3 ; sevE-sevP-stbm 7.1 /+ pk pk-sple-13 FRT42 pk pk-sple-13 cn sp/FRT42 pk pk-sple-13 cn pk pk-sple sev-pk sple w; FRT42 pk pk-sple-13 cn/FRT42 pk pk-sple-13 cn; sevE-sevP-sple 2.2 /+ w; stbm 6 /FRT42 P[w + ] stbm Vang-A3 w; FRT42 stbm 6 /FRT42 P[w + ] stbm Vang-A3 ; sevE-sevP-stbm 2.2 /+ w; FRT42 stbm 6 /FRT42 P[w + ] stbm Vang-A3 ; sevE-sevP-stbm 7.1 /+ FRT42 pk pk-sple-13 cn/FRT42 pk pk-sple-13 cn w; FRT42 pk pk-sple-13 cn/FRT42 pk pk-sple-13 cn; sevE-sevP-pk sple[2.2] /+ w; FRT42 pk pk-sple-13 cn/FRT42 pk pk-sple-13 cn; sevE-sevP-pk sple[14.2] /+ Figure 2 fz w; FRT42D/FRT42D Arm-fz-EGFP, Arm-lacZ; fz 15 /fz 21 , Ubx-FLP stbm/Vang y w Ubx-FLP/+; FRT42D stbm 6 /FRT42D Arm-lacZ stbm/Vang fz w; FRT42D stbm 6 /FRT42D Arm-fz-EGFP; fz 21 /fz 21 , Ubx-FLP stbm/Vang fz w; FRT42D stbm 6 , Arm-fz-EGFP/FRT42D Arm-lacZ; fz 21 /fz 21 , Ubx-FLP fmi/stan y w Ubx-FLP/+; FRT42D fmi E59 /FRT42D Arm-lacZ fmi/stan fz w; FRT42D fmi E59 /FRT42D Arm-fz-EGFP, Arm-lacZ; fz 21 /fz 21 , Ubx-FLP stbm/Vang fmi/stan y w Ubx-FLP/+; FRT42D stbm 6 fmi E59 /FRT42D Arm-lacZ Figure 3 dsh 3 y w dsh 3 FRT18A/w Arm-lacZ FRT18A; FLP38/+ dsh; fz y w dsh 3 f 36a FRT19A/w Arm-fz-EGFP FRT19A; fz 21 /fz 21 , Ubx-FLP dsh; stbm/Vang y w dsh 3 f 36a FRT19A/w Act-FRT-polyA-FRT-stbm-EYFP FRT19A; stbm 6 /stbm 6 , Ubx-FLP pk pk-sple y w Ubx-FLP/+; FRT42D pk pk-sple-13 /FRT42D Arm-lacZ pk pk-sple ; fz w; FRT42D pk pk-sple-13 /FRT42D Arm-fz-EGFP; fz 21 /fz 21 , Ubx-FLP pk pk-sple stbm/Vang y w Ubx-FLP/+; FRT42D pk pk-sple-13 stbm 6 /FRT42D Arm-lacZ Figure 4 stbm/Vang dgo y w Ubx-FLP/+; FRT42D stbm 6 dgo 380 /FRT42D Arm-lacZ pk pk-sple dgo y w Ubx-FLP/+; FRT42D pk pk-sple-13 dgo 380 /FRT42D Arm-lacZ pk pk-sple dgo; fz w; FRT42D pk pk-sple-13 dgo 380 /FRT42D Arm-fz-EGFP; fz 21 /fz 21 , Ubx-FLP Figure 5 stbm/Vang fz y w hsFLP1/+; Act-FRT-y + -FRT-GAL4, UAS-lacZ/+; fz 15 , UAS-stbm/Df(3L)fz D21 fz-EGFP stbm/Vang w hsFLP1/+; Act-FRT-y + -FRT-GAL4, stbm 6 /stbm 6 , UAS-fz-EGFP fz-EGFP pk pk-sple w hsFLP1/+; Act-FRT-y + -FRT-GAL4, pk pk-sple-13 /pk pk-sple-13 , UAS-fz-EGFP fz-EGFP pk pk-sple dgo 380 w hsFLP1/+; FRT42D pk pk-sple-13 dgo 380 /FRT42D pk pk-sple-13 dgo 380 ; Act-FRT-CD2-FRT-GAL4, UAS-fz-EGFP/+ fz-EGFP dsh 1 w dsh 1 /Y; FLP38/Act-FRT-y + -FRT-GAL4, UAS-lacZ; UAS-fz/+ fz-EGFP ptc-GAL4 dsh 3 y w dsh 3 f 36a FRT19A/y w w + FRT19A; ptc-GAL4/+; UAS-fz-EGFP, Ubx-FLP/+ Figure 6 Act-fz-EYFP pk pk-sple w hsFLP1/+; pk pk-sple-13 , Act-FRT-polyA-FRT-fz-EYFP/pk pk-sple-13 Act-fz-EYFP dsh 1 w dsh 1 /Y; Act-FRT-polyA-FRT-fz-EYFP/FLP38 dgo y w Ubx-FLP/+; FRT42D dgo 380 /FRT42D Arm-lacZ pk pk-sple y w Ubx-FLP/+; FRT42D pk pk-sple-13 /FRT42D Arm-lacZ pk pk-sple dgo y w Ubx-FLP/+; FRT42D pk pk-sple-13 dgo 380 /FRT42D Arm-lacZ dsh 3 y w dsh 3 FRT18A/w Arm-lacZ FRT18A; FLP38/+ Supplementary Figure 1 fz y w hsFLP1; Act-FRT-polyA-FRT-fz-EYFP/+; fz 21 fmi/stan y w hsFLP1; fmi E45 , GAL4-1407/fmi E59 ; Act-FRT-polyA-FRT-fmi-FLAG/UAS-fmi GAL4-1407 UAS-fmi fmi Usui et al., 1999 dsh w dsh 1 /Y; FLP38/+; Act>FRT-poly-FRT-dsh-ECFP/+ Supplementary Figure 2 w hsFLP1; stbm 6 /stbm Vang-A3 ; Act-FRT-polyA-FRT-stbm-EYFP/+ w hsFLP1; fmi E45 , GAL4-1407/fmi E59 ; Act-FRT-polyA-FRT-fmi-FLAG/UAS-fmi w dsh 1 /Y; FLP38/+; Act>FRT-poly-FRT-dsh-ECFP/+ w hsFLP1; pk pk-sple-13 /pk pk-sple-13 ; Act-FRT-polyA-FRT-pk/+ Supplementary Figure 3 stbm/Vang w hsFLP1; FRT42 stbm 6 /FRT42 P[w + ] stbm/Vang sev-stbm w hsFLP1; FRT42 stbm 6 /FRT42 P[w + ]; sevE-sevP-stbm 7.1 /+ fz 21 stbm 6 dsh 3 fmi E59 pk pk-sple-13 dgo 380 Jones et al., 1996; Wolff and Rubin, 1998; Wehrli and Tomlinson, 1998; Usui et al., 1999; Gubb et al., 1999; Feiguin et al., 2001 fmi E45 Usui et al., 1999 fz 15 Jones et al., 1996 stbm Vang-A3 Taylor et al., 1998 dsh 1 Perrimon and Mahowald, 1987; Axelrod et al., 1998; Boutros et al., 1998 Histology Strutt, 2001 Usui et al., 1999 Tree et al., 2002b Rawls and Wolff, 2003 Shimada et al., 2001 Feiguin et al., 2001 Tomlinson and Ready, 1987 Results Differing temporal requirements of the core polarity proteins during wing development fz fz fz Strutt and Strutt, 2002 fz fz ds Strutt and Strutt, 2002 fz stbm/Vang fz Taylor et al., 1998 fz Figs. 1 fz stbm/Vang Fig. 1 fz Supplementary Fig. 1A fz 21 Jones et al., 1996 stbm/Vang pk Tree et al., 2002b pk pk Figs. 1 stbm/Vang fz pk fmi/stan dsh Supplementary Figs. 1B, C dsh dsh 1 Perrimon and Mahowald, 1987 Feiguin et al., 2001 fmi/stan Supplementary Fig. 1B fz stbm/Vang dsh dsh Supplementary Fig. 1C fmi/stan fz stbm/Vang dsh dsh 1 Supplementary Fig. 2 Differing temporal requirements of the core polarity proteins during eye development fz stbm/Vang fz fz sevenless Strutt and Strutt, 2002 fz fz fz fz dsh 3 sev-dsh dsh fz stbm/Vang pk stbm/Vang sev-stbm Figs. 1 stbm/Vang fz pk sev-pk sple Figs. 1 Gubb et al., 1999 pk dsh stbm/Vang stbm/Vang stbm/Vang Supplementary Fig. 3 Taylor et al., 1998 stbm/Vang Wolff and Rubin, 1998 Supplementary Fig. 3 fz stbm/Vang fmi/stan fz stbm/Vang fmi/stan fz stbm/Vang fz Fig. 2 Gubb and García-Bellido, 1982; Vinson and Adler, 1987 stbm/Vang Fig. 2 Taylor et al., 1998 Strutt, 2001; Bastock et al., 2003 Lawrence et al., 2004 Amonlirdviman et al., 2005; Klein and Mlodzik, 2005; Le Garrec et al., 2006 fz stbm/Vang stbm/Vang; fz stbm/Vang stbm/Vang fz stbm/Vang fz stbm/Vang fz stbm/Vang fz fz stbm/Vang Lawrence et al., 2004 fz lacZ Fig. 2 stbm/Vang lacZ Fig. 2 stbm/Vang fz fz stbm/Vang Arm-fz-EGFP Strutt, 2001 stbm/Vang stbm 6 /stbm 6 ; fz 21 /fz 21 Arm-fz-EGFP/Arm-fz-EGFP; fz 21 /fz 21 Arm-fz-EGFP/stbm 6 ; fz 21 /fz 21 Materials and methods stbm/Vang; fz Fig. 2 Fig. 2 fz stbm/Vang Fig. 2 fz stbm/Vang Wong and Adler, 1993; Taylor et al., 1998 fz stbm/Vang fz + fz − stbm/Vang + stbm/Vang − Figs. 2 Usui et al., 1999; Bastock et al., 2003 Amonlirdviman et al., 2005; Klein and Mlodzik, 2005; Le Garrec et al., 2006 stbm/Vang; fz Fig. 2 fz stbm/Vang FRT42D stbm 6 , Arm-Fz-EGFP; fz 21 FRT42D Arm-lacZ; fz 21 Materials and methods stbm/Vang lacZ fz Arm-fz-EGFP lacZ stbm/Vang fz Fig. 2 Fig. 2 fz stbm/Vang Figs. 2 fz stbm/Vang Figs. 2 Fig. 1 stbm/Vang fz fz Figs. 2 stbm/Vang fz stbm/Vang fz Figs. 2 Wong and Adler, 1993; Taylor et al., 1998 fz stbm/Vang fmi/stan Fig. 2 Chae et al., 1999; Usui et al., 1999 fmi/stan Usui et al., 1999 fmi/stan fmi/stan Lawrence et al., 2004 fz fmi/stan stbm/Vang fmi/stan fmi/stan Figs. 2 fz Fig. 1 dsh and pk are not required for intercellular communication Tree et al., 2002b; Bastock et al., 2003; Jenny et al., 2005 dsh Fig. 3 Theisen et al., 1994 fmi/stan fmi/stan dsh dsh fz dsh stbm/Vang fz stbm/Vang Figs. 3 Fig. 3 dsh pk pk-sple pk Fig. 3 Gubb et al., 1999 pk-sple fz pk-sple stbm/Vang fz stbm/Vang Figs. 3 Fig. 3 pk fz stbm/Vang Vinson and Adler, 1987; Taylor et al., 1998; Adler et al., 2000 dsh pk Analysis of dgo function Drosophila dgo stbm/Vang pk Das et al., 2004 pk dgo stbm/Vang dgo stbm/Vang Fig. 4 pk dgo pk Fig. 4 pk dgo fz fz Fig. 4 dgo pk stbm/Vang Das et al., 2004 stbm/Vang dgo Fig. 4 stbm/Vang Bastock et al., 2003 pk dgo Fig. 4 pk dgo Fig. 4 pk pk dgo dsh fz Lawrence et al., 2004 Amonlirdviman et al., 2005; Klein and Mlodzik, 2005; Le Garrec et al., 2006 dsh pk dgo fz pk Lawrence et al., 2004 pk 1 pk Adler et al., 2000 pk fz stbm/Vang Strutt, 2001 Amonlirdviman et al., 2005 fz stbm/Vang Taylor et al., 1998 fz Fig. 5 stbm/Vang Fig. 5 pk Fig. 5 n Adler et al., 2000; Lawrence et al., 2004 pk dgo Fig. 5 dgo pk dsh dsh dsh 1 Fig. 5 n dsh patched Adler et al., 1997 dsh 3 Fig. 5 Wong and Adler, 1993 Asymmetry of polarity protein activity exists in the absence of asymmetry in distribution Amonlirdviman et al., 2005; Klein and Mlodzik, 2005; Le Garrec et al., 2006 pk dsh 1 Figs. 5 Adler et al., 2000; Lawrence et al., 2004 Figs. 6 Axelrod, 2001; Strutt, 2001; Usui et al., 1999 Strutt, 2001 pk dsh 1 Figs. 6 pk dsh 1 dgo pk Figs. 6 dgo pk Fig. 6 pk dgo dsh fz stbm/Vang Fig. 6 Wong and Adler, 1993; Taylor et al., 1998 dsh pk dgo Discussion Introduction (i) The transmembrane proteins Fz, Stbm/Vang and Fmi/Stan have a common early function during planar polarity patterning in the wing and eye, with the cytoplasmic factors Dsh and Pk playing only later roles. (ii) and (iii) Intercellular communication does not require Dsh or Pk in either Fz or Stbm/Vang expressing cells and Dgo also does not play redundant roles with Pk in intercellular signalling. (iv) Pk and Dgo act redundantly in propagation of polarity from cell to cell, most likely by promoting intracellular communication. Dsh plays a prominent role in such propagation, greater than that of both Pk and Dgo. We speculate that the intracellular communication required for such polarity propagation is dependent on establishing intracellular asymmetries of protein activity. (v) pk (vi) Neither Pk nor Dgo are directly required for determining the site of trichome placement. Coupling to the global polarity cue Introduction fj/ds/ft Ma et al., 2003; Yang et al., 2002 fz stbm/Vang fmi/stan ds Matakatsu and Blair, 2004 pk dsh pk Lawrence et al., 2004 pk Local coordination of polarity Lawrence et al., 2004 Amonlirdviman et al., 2005; Le Garrec et al., 2006 Amonlirdviman et al., 2005; Klein and Mlodzik, 2005; Le Garrec et al., 2006 pk dsh 1 Lawrence et al., 2004 fz stbm/Vang dsh pk dgo pk Adler et al., 2000; Lawrence et al., 2004 pk dgo Lawrence et al., 2004 Amonlirdviman et al., 2005; Klein and Mlodzik, 2005; Le Garrec et al., 2006 Amonlirdviman et al., 2005 Le Garrec et al., 2006 pk dgo pk dsh 1 pk Amonlirdviman et al., 2005 Ma et al., 2003; Amonlirdviman et al., 2005 pk Classen et al., 2005 Specification of the site of polarised trichome production Adler et al., 2004 Concluding remarks Lawrence et al., 2004; Amonlirdviman et al., 2005; Klein and Mlodzik, 2005; Le Garrec et al., 2006 Appendix A Supplementary data Supplementary Fig. 1 fz fmi/stan dsh 1 Act fz-EYFP fz 21 fz fz Strutt and Strutt, 2002 fz Act fmi-FLAG fmi E45 /fmi E59 fmi/stan fmi/stan UAS-Fmi Usui et al., 1999 fmi/stan fz fz Act dsh-ECFP dsh 1 dsh fz stbm/Vang fmi/stan  Supplementary Fig. 2 stbm-EYFP stbm 6 /stbm Vang-A3 fmi-FLAG fmi E45 /fmi E59 dsh-ECFP dsh 1 pk pk pk pk-sple-13  Supplementary Fig. 3 stbm/Vang stbm 6 stbm 6 stbm/Vang stbm/Vang stbm 6 stbm 15 stbm/Vang stbm 6 stbm 15 stbm/Vang stbm/Vang stbm/Vang Wolff and Rubin, 1998 stbm/Vang Taylor et al., 1998 stbm/Vang Wolff and Rubin, 1998 stbm/Vang stbm/Vang Taylor et al., 1998; Adler et al., 2000 stbm/Vang stbm/Vang Wolff and Rubin, 1998 stbm 6 stbm sevenless stbm sevenless stbm/Vang stbm/Vang  Appendix A Supplementary data doi:10.1016/j.ydbio.2006.09.026