spacer gif spacer gif spacer gif spacer gif spacer gif
QUICK SEARCH:   [advanced]


spacer gif
     Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents    

First published online August 4, 2003
doi: 10.1242/10.1242/dev.00639

This Article
Right arrow Summary Freely available
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Jenkins, A. B.
Right arrow Articles by Van Doren, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Jenkins, A. B.
Right arrow Articles by Van Doren, M.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati   Add to Twitter  
What's this?

Drosophila E-cadherin is essential for proper germ cell-soma interaction during gonad morphogenesis

Allison B. Jenkins1, J. Michael McCaffery2 and Mark Van Doren1,*

1 Department of Biology, Mudd Hall, Johns Hopkins University, 3400 N. Charles Street, Baltimore, MD 21218, USA
2 Integrated Imaging Center, Department of Biology, Mudd Hall, Johns Hopkins University, 3400 N. Charles Street, Baltimore, MD 21218, USA



View larger version (124K):

[in a new window]
  		Fig. 3. Drosophila E-cadherin is expressed in germ cells and SGPs during gonad coalescence. Anterior is leftwards. All embryos are stained for Drosophila E-cadherin. (A-D) Expression of E-cadherin in stage 12 (A), stage 13 (B), stage 14 (C) and stage 17 (D) gonads. (C,D) Male embryos. SGPs and msSGPs (large arrow in C) are labeled with anti-EYA in A-C. Gonad boundaries (SGPs) indicated with broken lines. E-cadherin expression in the SGPs appears between stages 12 (A) and 13 (B), correlating with the onset of coalescence. E-cadherin expression is maintained in the gonad throughout embryogenesis and is increased at the anterior of the gonad during late stages (a, anterior; D). See text for further discussion of symbols. (E) Stage 15 embryo double labeled for E-cadherin and Armadillo/ß-catenin. Note the extensive co-localization of these proteins. (F) Stage 15 male. UAS-mCD8-GFP expressed in germ cells (nos-Gal4) co-localizes with E-cadherin, indicated by yellow, except in regions of germ cell-germ cell contact (black arrowhead). msSGPs (arrow) also exhibit E-cadherin expression. Higher magnification view of two germ cells co-expressing mCD8-GFP and E-cadherin on their cell surfaces. There is also a region of E-cadherin alone between these two cells that is probably an ensheathing SGP. (G) Stage 11. E-cadherin is present at the surface of migrating germ cells (asterisks). Germ cells were identified by co-expression of mCD8-GFP crossed to nanos-Gal4 (not shown). (H) Stage 15. mCD8-GFP expressed in the mesoderm (24B-Gal4) co-localizes with E-cadherin. (I) Stage 15 agametic (osk) embryo, EYA labels SGPs. Note that E-cadherin is expressed even though no germ cells are present. (J) Stage 14 eyaCli-IID mutant. Germ cells labeled with Vasa clearly express E-cadherin. Surrounding ZFH-1-positive cells, potentially SGPs or other mesodermal cells, do not display E-cadherin staining. t, trachea; hg, hindgut. Scale bar in A: 10 µm.

 


View larger version (103K):

[in a new window]
  		Fig. 2. TEM analysis of the coalesced gonad. (A,B) Stage 14 male wild type gonad, transverse section. Two germ cells are highlighted in yellow, two SGPs highlighted in pink and purple. (A) SGPs ensheath germ cells with thin cellular extensions. Note the unusual morphology of the purple SGP as it extends processes to ensheath the neighboring germ cell. (B) Higher magnification of region indicated in A. Two SGPs overlap each other to completely surround a germ cell.

 


View larger version (133K):

[in a new window]
  		Fig. 1. SGPs individually ensheath germ cells during gonad formation. Anterior is leftwards. (A-E) UAS-mCD8-GFP expressed in the germ cells (nos-Gal4; A,B) or the mesoderm (24B-Gal4; C-E) of stage 13 (A,C), stage 15 (B,D) or stage 16 (E) embryos. (A,B) Anti-GFP labels the surface of germ cells, SGPs are labeled with anti-EYA. Note that the germ cells are round during gonad coalescence. (B, inset) Migrating germ cell (stage 11). (C,D) Somatic cell surfaces labeled with anti-GFP and SGP nuclei labeled with anti-EYA. Embryos are male, identified by the presence of EYA-positive msSGPs at the posterior of the gonad. Germ cells (e.g. asterisk) are surrounded by thin, SGP-derived cellular extensions in the uncoalesced (C) and coalesced (D) gonad. (E) Somatic cell surfaces labeled with anti-GFP and germ cells labeled with anti-Vasa. Closely apposed germ cells remain extensively ensheathed in late stages of embryogenesis. (F) Agametic stage 15 embryo (osk). SGPs labeled with anti-ZFH-1; anti-Neurotactin (NRT) labels the surface of many cell types, including the SGPs. Broken line indicates the boundary of the gonad. Arrows indicate SGP extensions around germ cells. Scale bar in A: 10 µm.

 


View larger version (79K):

[in a new window]
  		Fig. 4. shg/E-cadherin mutants have defects in gonad compaction, germ cell ensheathment and germ cell migration. Anterior is leftwards, embryos are stage 15 or later. Vasa labels the germ cells in B-D,F. (A,B) shgG317 homozygous embryos. (A) SGPs labeled with EYA. Example of a strong gonad compaction defect. shg mutants also display defects in germ cell migration, and the gonad in A lacks germ cells completely (Vasa channel not shown). (B) EYA marks the SGPs. Example of a weak gonad compaction defect. (C) Stage 15 wild-type male. msSGPs (arrow) express EYA and Sox100B, while SGPs express only EYA. msSGPs join the posterior of male gonads. (D) shgG317 homozygous male embryo. msSGPs have failed to join the gonad, remaining in a tight cluster posterior to the gonad (arrow). (E) shgG317 homozygous embryo. UAS-mCD8-GFP expressed in the mesoderm (24B-Gal4) labeled with anti-GFP and anti-EYA. The lack of GFP-labeled SGP extensions between the germ cells indicates a failure of germ cell ensheathment. (F) Stacked z-series through an embryo from a cross of shgG317/CyOftz-lacZ females and w1118 males that did not inherit the shgG317 chromosome zygotically. Anti-EYA (not shown) was used to identify the normal position of the one gonad visible in this image (circle). Many lost germ cells are observed, indicating a dominant maternal effect of shgG317 on germ cell migration. Scale bar in A: 10 µm.

 


View larger version (131K):

[in a new window]
  		Fig. 5. A balance of E-cadherin is crucial for normal gonad formation. Anterior is leftwards. Stage 15 embryos carrying an enhancer trap (68-77) that expresses ß-GAL in SGPs. Embryos are labeled to reveal germ cells (anti-Vasa), the enhancer trap (anti-ß-GAL) and the cell surface of SGPs (anti-NRT). (A,A') Wild type. Germ cells are completely ensheathed by SGPs. Note that both ß-GAL and NRT staining is observed between germ cells. (A') NRT channel alone. (B,B') Embryo expressing two copies of a UAS-DE-cadherin transgene in the germ cells (nos-Gal4). Germ cells are tightly clustered inside the gonad, with no ß-GAL or NRT labeling between them. (B') NRT channel alone. Scale bar in A: 10 µm.

 


View larger version (104K):

[in a new window]
  		Fig. 6. foi is required for proper Drosophila E-cadherin levels in the gonad. Anterior is leftwards. (A) Stage 15 foi20.71/foi16.33 mutant. UAS-mCD8-GFP expressed in the mesoderm (twist-Gal4) labeled with anti-Vasa, anti-GFP and anti-NRT. The absence of SGP-derived GFP and NRT labeling around the germ cells indicates a failure in germ cell ensheathment. (B,C) foi20.71 homozygous mutants. Embryos labeled with anti-Vasa, anti-EYA and anti-Drosophila E-cadherin. Gonad boundaries indicated with broken line. (B) Drosophila E-cadherin staining is reduced in the SGPs of foi mutants at stage 13 compared with wild type (Fig. 3). (C) Stage 15 male. The SGPs and germ cells have not compacted into a coalesced gonad but the msSGPs (C, arrow) have joined the gonad. Drosophila E-cadherin expression remains low in the SGPs and germ cells, but is high in the msSGPs (arrow). Nearby tissues (t, trachea) are not affected. Scale bar in A: 10 µm.

 

Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati   Add to Twitter Twitter    What's this?



© The Company of Biologists Ltd 2003