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doi: 10.1242/10.1242/dev.00574

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PVF1, a PDGF/VEGF homolog, is sufficient to guide border cells and interacts genetically with Taiman

Jocelyn A. McDonald*, Elaine M. Pinheiro* and Denise J. Montell{ddagger}

Department of Biological Chemistry, The Johns Hopkins University School of Medicine, 725 North Wolfe Street, Baltimore, MD 21205-2185, USA



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  		Fig. 1. Genetic screen to identify dominant interactions with tai. (A) Nomarski optics image of an ovariole stained for ß-galactosidase activity from the enhancer trap PZ8685, showing the germarium stage (far left) to stage 10 (S10; far right). PZ8685 is detected primarily in the border cells (bc, arrows), which migrate at stage 9 (S9) through the nurse cell cluster (nc). The border cells complete their migration at stage 10, when they reach the anterior border of the oocyte. (B,C) Representative examples of border cell (arrows) migration defects observed in stage 10 egg chambers heterozygous for tai61G1 (tai) and either Df(1)N19 (B) or Df(3)GN24 (C). (D) Crossing scheme used for the tai genetic interaction screen. The arrowheads in A-C indicate the extent of rearrangement of the outer follicle cells (ofc). Anterior is towards the left in all panels. Df, deficiency.

 


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  		Fig. 2. All deficiencies screened for dominant interaction with tai. The Bloomington Stock Center deficiency kit was screened for dominant genetic interaction with tai, indicated by the boxes below each chromosome arm (thick black lines). The identity of the chromosome arms are indicated on the left. The size of the box corresponds to the cytological breakpoints for the deficiency, indicated by the numbers below the chromosome arms. White boxes represent deficiencies that were tested but had no interaction with tai. Black boxes represent deficiencies that interacted with tai (Table 1). The gray box represents the deficiency that removes the tai locus at the 30A6 region.

 


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  		Fig. 3. tai exhibits dominant genetic interactions with Pvf1. (A) PZ8685 staining in a tai61G1/CyO egg chamber at stage 10. The border cells (arrow) have completed their migration and reside at the anterior border of the oocyte (arrowheads). The dashed line indicates the distance 50% along the length of the migration route. The dotted line indicates the distance 90% along the length of the migration route. The border cells in this egg chamber have migrated more than 90% of the normal distance. (B) PZ6356 staining, which labels the border cells, the oocyte nucleus and some nurse cell nuclei, in a Pvf1EP1624/+; tai61G1/+ double heterozygous egg chamber at stage 10. Border cell migration (arrow) is delayed. In this egg chamber, the border cells have migrated ~60% of the normal distance. (C,D) Quantitation of border cell migration shown as the proportion of egg chambers in which the border cells migrated 0-25% (yellow), 26-50% (blue), 51-75% (red) and 76-100% (black) of the normal distance. Border cell migration was assayed in the indicated genotypes at stage 10. The number of egg chambers examined is indicated (n). (C) Interaction of tai with Pvf1EP1624 (EP1624). (D) Interaction of tai with slbo-GAL4/UAS-Pvf1.

 


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  		Fig. 4. E-cadherin distribution and TAI and PVR expression in mutant egg chambers. (A,B) Expression of TAI in wild-type (A) and Pvf1EP1624 (B) egg chambers. Border cells are indicated by the arrow. (C,D) PVR expression (red) in the follicle cells surrounding the oocyte in an egg chamber containing a large tai mutant follicle cell clone. Absence of GFP staining (green in D) marks the cells that are homozygous mutant for tai61G1 (tai-/-; broken line indicates the clone border). (E,F) PVR expression in wild-type (E) and tai mutant (F) border cell clusters (arrows). (G-L) Merged confocal z sections showing DE-cadherin protein expression in border cell clusters. (G) An example of a stage 9 wild-type border cell cluster undergoing migration. The polar cells (*) express highest levels of DE-cadherin, whereas the border-cell/nurse-cell junctions exhibit lower, punctate staining for DE-cadherin (arrowheads). DE-cadherin levels are high at junctions between border cells. (H) Stage 10 tai mutant border cell cluster. DE-cadherin protein levels are high throughout the cluster (compare with polar cells, *), especially at several nurse-cell/border-cell boundaries (arrows). (I,J) Stage 10 Pvf1 mutant egg chamber. DE-cadherin levels are high at a subset of border-cell/nurse-cell junctions (arrows). (J) DE-cadherin is high between border cells (compare with polar cells, *) and in one border-cell/nurse-cell junction. (K,L) Stage 10 slbo-GAL4/UAS-Pvf1 (UAS-Pvf1) border cell clusters. DE-cadherin levels are high at border-cell/nurse-cell junctions (arrows); polar cells are marked with an asterisk. Anterior is towards the left in all panels.

 


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  		Fig. 5. PVR function in border cell migration. (A,B) Wild-type expression of PVF1 (green) in the oocyte and nurse cell cytoplasm and PVR (red) in follicle cells at stage 8 (A) and stage 9 (B). PVR is expressed in the border cells (arrows). (C,D) PVR expression (red) in the outer follicle cells of an egg chamber with several Pvrc2195 mosaic clones. Loss of GFP (green in D) marks cells homozygous mutant for Pvr (broken lines surround the mutant clones). (E,F) Egg chambers with border cells (arrows) mutant for Pvr. GFP (green) positively marks the homozygous mutant cells and filamentous actin (red) outlines all cells. (E) Egg chamber in which the mutant border cell cluster (arrow) exhibited delayed migration. (F) Egg chamber in which the mutant border cell cluster (arrow) migrated to the oocyte. (G) Quantitation of the Pvr mutant migration defect compared with the Pvf1 migration defect. Quantitation of border cell migration shown as the proportion of egg chambers in which the border cells migrated 0-25% (yellow), 26-50% (blue), 51-75% (red) and 76-100% (black) of the normal distance. Anterior is towards the left in all panels.

 


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  		Fig. 6. (A) Wild-type expression of PVF1 (red) in the oocyte and nurse cell cytoplasm at stage 9. (B,C) Clones of ectopic PVF1 expression in follicle cells of a stage 9 egg chamber showing expression of PVF1 in red (B) co-expressed with ß-galactosidase (green), which labels the clones (C). The broken lines show the outline of the egg chambers. (D-G) Classes of phenotypes observed after misexpression of PVF1, PVF2 or GRK. Egg chambers with clones of anterior follicle cells misexpressing one or more ligands (green, arrowheads). Singed staining labels the border cells (red, arrows). Representative examples of the four phenotypic classes are shown. (D) Class I: normal migration of the border cell cluster; an example of a UAS-Pvf1 egg chamber is shown. (E) Class II: the border cells fail to migrate away from the anterior pole and are adjacent to a clone of follicle cells misexpressing ligand; an example of a UAS-Pvf1 egg chamber is shown. (F) Class III: the border cells localize to the side of the egg chamber less than one nurse cell away from the anterior pole and adjacent to follicle cells misexpressing ligand; an example of a UAS-Pvf1;UAS-grk egg chamber is shown. (G) Class IV: the border cell cluster localizes to the side of the egg chamber at least two nurse cells away from the anterior pole, adjacent to a clone of cells misexpressing ligand; an example of a Pvf11624;UAS-Pvf1 egg chamber is shown. (Insets) Schematics of the stage 10 egg chambers represented in the fluorescent images, depicting the position of the border cells (red) with respect to the cells misexpressing ligand (green). The area indicated by the blue dashed box in the schematic outlines the anterior half of the egg chamber, which is shown at high magnification in the fluorescent images. Anterior is towards the left in all panels.

 

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© The Company of Biologists Ltd 2003