Fig. 2. VVL expression pattern in lch5 organs and rescue experiments. Anterior is towards the left and dorsal is upwards in all panels. (A-F) ato-lacZ embryos stained with anti-VVL (green) and anti-ß-GAL (red). (A) An early stage 11 embryo showing VVL expression in the tracheal placodes (t). VVL is not expressed in the ch precursors. (B,C) Abdominal segments of a stage 12 embryo. The box in B marks VVL-positive cells that belong to the lch5 lineage (arrow) and the oenocytes (arrowheads). (C) A merge of anti-VVL (B) and anti-ß-GAL staining. VVL-positive nuclei boxed in B colocalize with the ato-lacZ staining (arrows). The oenocytes are marked with arrowheads. (D-F) A stage 15 embryo. VVL expression is clearly detected in the nuclei of lch5 neurons (white box in D and arrows in F). (G-J) ß-GAL expression under the regulation of arm-Gal4 (G,H), ato-Gal4 (I) and elav-Gal4 (J). (G,H) Two focal planes of the same stage 14 embryo. arm-Gal4 drives strong ß-GAL expression (red) in the ectoderm (G), and much weaker expression in the PNS (H). Ectodermal and PNS focal planes were determined by staining with anti-85E (green). The arrow in G indicates lch5 ectodermal attachment cells. The arrow in H indicates the position of neurons and sheath cells. c, cap cells; l, ligament cells. (I) ato-Gal4 drives expression in the ch lineage and in a group of ectodermal cells that remains confined to the posterior compartment of each segment. lch5 in the A1 segment are boxed. White arrows in the T3 segment indicate the edges of dch3 organs, and black arrows indicate the cluster of the ß-GAL-expressing ectodermal cells. (J) lacZ expression induced by elav-Gal4 can be observed in the neurons (n) and sheath cells (s) of lch5 organs. (K-M) Rescue experiments with elav-Gal4 and arm-Gal4. (K,L) Embryos are labeled with mAb 22C10 (red) and anti-VVL (green). (K) A normal embryo carrying both the elav-Gal4 and UAS-vvl transgenes. The lch5 organs are positioned correctly in the lateral clusters in all abdominal segments (arrow). VVL expression is seen in the ectoderm and trachea (endogenous protein), and in neurons and sheath cells (induced protein). (L) A ut^H599 homozygous embryo, carrying both transgenes as the embryo in K. VVL expression can be observed only in neurons and sheath cells. This expression is clearly insufficient for rescuing the abnormal localization of lch5 organs. Abnormally positioned lch5 neurons are marked with arrowheads. (M) Abdominal segments of a ut^H599 homozygous embryo, carrying the arm-Gal4 and UAS-vvl transgenes. Note the correct positioning of lch5 neurons. D, dorsal PNS cluster; L, lateral PNS cluster.

Return to article